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PL0001643

POLISH ACADEMY OF SCIENCES INSTITUTE OF BIOCHEMISTRY AND BIOPHYSICS RESEARCH REPORT

1996-1997 INSTITUTE OF BIOCHEMISTRY AND BIOPHYSICS Polish Academy of Sciences

Director: Wlodzimierz Zagorski-Ostoja

Head of the Scientific Council: Andrzej Paszewski (till 22.08.1997 Zofia Lassota)

Deputy Scientific Director: Piotr Ceglowski

ADDRESS, E-MAIL, FAX AND TELEPHONE:

5a Pawinskiego Street, 02-106 Warsaw, Poland

E-Mail: [email protected] International telephone and Fax network: (48) 39-12-16-23 Domestic telephone: (48-22) 658-44-99, (48-22) 659-70-72 Domestic Fax: (48-22) 658-46-36 Switchboard: (48-22) 658-44-99, (48-22) 659-70-72 Director / Secretariate: (48-22) 658-47-24; Fax: (48-22) 658-46-36 Scientific Council: (48-22) 658-44-99 ext. 24-12 or 21-40 Scientific Secretariate: (48-22) 658-47-53 Polish-French Center of Plant Biotechnology:(48-22) 658-46-60 School of Molecular Biology: (48-22) 658-47-02 Warsaw University Affiliates: Department of Genetics: (48-22) 658-47-54 Institute of Experimental Plant Biology: (48-22) 658-39-49 fax: (48-22) 658-48-04 Administration: (48-22) 658-47-00 Guest rooms: (48-22) 658-47-94

ISBN 83-906782-1-7 The Institute of Biochemistry and Biophysics of the Polish Academy of Sciences was founded in Warsaw in 1957. Initially a biochemical research center, the Institute presently specializes in advanced molecular biology of plants, fungi and microorganisms and in state-of-the-art computer modeling and bioinformatics.

Specialized areas of interest include:

gene sequencing and mapping regulation of gene expression at DNA and RNA levels protein biosynthesis and post-translational modification mutagenesis and DNA repair metabolism of nucleic acids protein kinases regulation of enzyme activity protein structure-function studies computer modeling of peptides and proteins NMR studies of proteins and peptides antiviral and anticancer nucleotides polyprenoids structure and function nucleus-mitochondria interaction molecular plant virology plant transformation molecular basis of plant and microbial biotechnology CONTENTS

Institute profile 5 Scientific Council 9 Trustee Council 13 School of Molecular Biology 15 Polish-French Center of Plant Biotechnology 16 International Collaboration 17 Library 18

Research Units of the Institute Department of Biophysics 21 Department of Genetics 28 Department of Lipid Biochemistry 33 Department of Microbial Biochemistry 36 Department of Molecular Biology 42 Department of Plant Biochemistry , 48 Department of Protein Biosynthesis 53 Laboratory of Mutagenesis and DNA Repair 59 Laboratory of Purine Metabolism 62 NMR Laboratory 63 Bioinformatics 64

Laboratory of Molecular Biology 65 (affiliated with the University of Gdansk)

Research Units of the Warsaw University located at the Institute Department of Genetics (associated with the Institute) 69 Institute of Experimental Plant Biology 71

Research grants , 76

Meetings, Symposia, Courses & Seminars 83

List of Publications Publications , 92 Abstracts. Miscellaneous : 112 Patents 140 Dissertations ...V. 141

Institute Staff Members 148 Several research and educational organizations associated with the Institute are located in newly constructed quarters of IBB with over 12,000 square meters of laboratory space

They include:

IBB Polish-French Center for Plant Biotechnology Research sponsored by the Governments of both countries

Interdisciplinary Center for Advanced Computer Modeling, Warsaw University highly professional staff and modern facility equipped with Cray computers

Institute of Experimental Plant Biology and Departament of Genetics Warsaw University which complement IBB expertise

IBB Post-graduate School of Molecular Biology with over 70 students working towards the Ph. D. degree

IBB NMR (500 MHz) Laboratory

IBB Polish National Node of European Molecular Biology Network (EMBNet) which provides access to major gene banks and related programs and information for over 600 scientists in Poland

Regional Node of International Center for Cooperation in Bioinformatics (ICCB) under the auspices of UNESCO Institute Profile Organization and Activities

The Institute of Biochemistry & Biophysics (IBB) of the Polish Academy of Sciences (PAS; PAN in Polish) was founded in 1957. But it was only in 1994-1995 that it was able to move into its own, appropriately designed, new buildings on Pawinskiego Street, with a floor space of 12,500 sq. m., equipped with modern laboratories, computer systems, and a telecommunication network in line with world standards.

The staff of the Institute currently comprises 260 persons, of whom 186 are involved in research, and the remainder in services and administration. The various Departments and independent Laboratories, along with their research and other activities, are listed in full detail below.

The Institute is administered by a Director, supported by a Board (consisting of Heads of departments, laboratories and research teams) and an advisory Scientific Council, the membership of which includes scientists from other centers in Poland (see below for list of members). There is also a Trustee Council, consisting of former members of the Institute who now reside permanently abroad at various academic or other research centers, but who desire to maintain close scientific contacts with the Institute (see below for list of members).

In general, scientific interests of the Institute have evolved over the years from classical biochemistry, biophysics and physiological chemistry to up-to-date molecular biology. Research interests are focussed on replication, mutagenesis and repair of DNA; regulation of gene expression at various levels; biosynthesis and post-translational modifications of proteins; gene sequencing and functional analysis of open reading frames; structure, function and regulation of enzymes; conformation of proteins and peptides; modeling of structures, and prediction of functions of proteins; mechanisms of electron transfer in polypeptides. Although basic research is considered the primary function, attention of several research groups is simultaneously oriented to potential clinical and phytopathological applications.

IBB has, from its inception, maintained close contacts with centers of higher education, both in research and teaching. A good example is the Department of Biophysics of the Institute of Experimental Physics of the Warsaw University, initially organized and directed by staff members of IBB, headed by Prof. D. Shugar, and close collaboration between our Institute and the Department has continued unabated over the years.

The Institute is formally and closely associated with the Faculty of Biology of the Warsaw University, leading to the location on the site of our Institute of the Department of Genetics and the Institute of Experimental Plant Biology of the University. Recently a division of the Institute was created at the University of Gdansk. These affiliations with university centers have contributed not only to an expansion of the research actvities of the Institute, but also to a program of advanced education in molecular biology through the IBB School of Molecular Biology, with more than 70 young researchers enrolled for the Ph.D. degree (see report, below, on the School of Molecular Biology). It should be noted here that IBB possesses the necessary authorization to award higher degrees (Ph.D., Dr.Sc.) in the fields of biochemistry, biophysics and genetics.

Educational activities of the Institute concern specialized courses which are organized in collaboration with the Departments from the Faculty of Biology of the Warsaw University. The courses include training in basic methods of molecular biology and biotechnology, e.g. gene cloning and sequencing, analysis of gene expression etc., with special emphasis on bioinformatics. These educational activities are partly supported by the special PHARE SCI-TECH Project.

Functional activities of the Institute include Bioinformatics. Current releases of major molecular biology databases, updated daily, are available on the Institute network, along with tools for software analysis. Our internal structural network permits access to software and data maintained on a multiprocessor Silicon Graphics system from more than 100 computers and terminals available throughout the Institute. The Institute internal network is accessible country-wide through Internet, and over 600 molecular biology database users are registered at the Institute mainframe. The quality of our bioinformatics setup has led to the recognition of IBB as the national node of the European Molecular Biology Network, so that IBB is considered a unique bioinformatics facility in Poland. Bioinformatics is also being pursued within the framework of a Poland-Israel program, supported by UNESCO, and codirected by IBB and the Weizmann Institute of Science in Rehovot.

A recent result of this cooperation is that since April 1997 the Institute acts as a Regional Node of UNESCO ICCBnet - a network for promoting bioinformatics to biotechnologically developing countries. IBB has undertaken the responsibility of the regional node in Central and Eastern Europe. ICCBnet has been approved as the official UNESCO network by the resolution of the General Conference in November 1997.

The Institute has at its disposal several funds for the expansion and promotion of research activities:

(a) Stipends for candidates formally registered in study programs leading to a higher degree.

(b) Repatriation fund, in the form of financial assistance to former members of the Institute, presently working in academic or other research centers outside Poland, and who have expressed a desire to return to IBB. (c) European Fellowship Fund: This Fund offers fellowships to scientists from other research centers in Central and Eastern Europe, enabling them to spend periods of 3 to 6 months at our Institute or affiliated Polish scientific centers, either on a pre- determined research project, or on one already being conducted in collaboration with the candidate's parent institute. This is facilitated by the existence, in our new buildings, of several comfortable guest rooms, also available for invited lecturers.

Collaboration with foreign laboratories, initiated from the period of inception of the Institute, has been particularly accentuated in the past five years, especially with centers in France. In 1991 IBB became formally associated with the Centre de Genetique Moleculaire (CGM) of the Centre National de la Recherche Scientifique (CNRS), and the Institut Jacques Monod of the Universite de Paris VII. The Polish- French Center of Plant Biotechnology (see below for further details) was established in 1994 by an accord between the CNRS, the Polish Academy of Sciences, and the Polish State Committee for Scientific Research (KBN). This has already enabled many members of our staff to spend short-term (1-3 months) working visits in French molecular biology laboratories, leading to the several collaborative research projects, while many French scientists have visited our Institute, mainly to deliver lectures and seminars to our Ph.D. students in the School of Molecular Biology. Further details of international collaborative projects are outlined below.

NEXT PAGE(S) left BLANK SCIENTIFIC COUNCIL 1996- 1998

Members of the Polish Academy of Sciences:

Tadeusz CHOJNACKI Corresponding Member, Institute of Biochemistry and Biophysics, PAS ul. Pawinskiego 5a, 02-106 Warszawa

| Waclaw GAJEWSKI | Member, Institute of Biochemistry and Biophysics, PAS ul. Pawinskiego 5a, 02-106 Warszawa

Wlodzimierz OSTROWSKI Member, Jagiellonian University, Collegium Medicum, Institute of Medical Biochemistry ul. Kopernika 7, 31-034 Krakow

David SHUGAR Foreign Member, Institute of Biochemistry and Biophysics, PAS ul. Pawinskiego 5a, 02-106 Warszawa

Piotr P. SLONIMSKI Foreign Member, Centre de Genetique Moleculaire, CNRS 91190 Gif-sur-Yvette, France

Przemyslaw SZAFRANSKI Member, Institute of Biochemistry and Biophysics, PAS ul. Pawinskiego 5a, 02-106 Warszawa

| Karol TAYLOR | Corresponding Member, University of Gdansk, Department of Molecular Biology ul. Kladki 24, 00-822 Gdansk

Piotr W^GLENSKI Corresponding Member, Warsaw University, Department of Genetics/Institute of Biochemistry and Biophysics PAS, ul. Pawinskiego 5a, 02-106 Warszawa Kazimierz L. WIERZCHOWSKI Member, Institute of Biochemistry and Biophysics, PAS ul. Pawiriskiego 5a, 02-106 Warszawa

Lech WOJTCZAK Member, Institute of Experimental Biology, PAS ul. Pasteura 3, 02-095 Warszawa

Members of the Council from outside of the Institute

Ryszard W. ADAMIAK Institute of Bioorganic Chemistry, PAS ul. Noskowskiego 12/16, 60-637 Poznari

Jan ALBRECHT Center of Experimental and Clinical Medicine, PAS ul. Pawinskiego 5, 02-106 Warszawa

Jerzy DUSZYNSK1 Institute of Experimental Biology, PAS ul. Pasteura 3, 02-095 Warszawa

Witold FILIPOWICZ Friedrich-Miescher-Institut CH-4002 Basel, Postfach 2543, Switzerland

Marek GNIAZDOWSKI Medical Academy ul. Lindleya 6, 90-131 Lodz

Marian KOCHMAN Wroclaw Polytechnical University ul. Wybrzeze Wyspianskiego 27, 51-114 Wroclaw

Liliana KONARSKA Medical Academy, Warsaw, Institute of Biopharmacy ul. Banacha 1, 02-097 Warszawa

Bogdan LESYNG Warsaw University, ICM ul. Pawinskiego 5a, 02-106 Warszawa

10 Stefan MALEPSZY Agricultural Academy, Warsaw, Department of Genetics and Plant Breeding ul. Nowoursynowska 166, 02-789 Warszawa

Jacek OTLEWSKI University of Wroclaw, Faculty of Natural Sciences, Institute of Biochemistry ul. Tamka 1, 50-137 Wroclaw

Andrzej PLUCIENMCZAK Institute of Antibiotics and Biotechnology ul. Staroscinska 5, 02-216 Warszawa

Leon SEDLACZEK Center of Microbiology and Virology, PAS ul. Lodowa 106, 93-232 Lodz

KrzysztofSTARON Faculty of Biology, Department of Biochemistry, Warsaw University ul. Zwirki i Wigury 93, 02-089 Warszawa

Zygmunt WASYLEWSKI Institute of Molecular Biology, Jagiellonian University Al. A. Mickiewicza 3, 31-120 Krakow

Staff Members of the Institute:

Ewa BARTNIK (Warsaw University/IBB PAS) Andrzej BIERZYNSKI Jerzy BUCHOWICZ Piotr CEGLOWSKI Zygmunt CIESLA Magdalena FIKUS Danuta M. HULANICKA-DZIECHCINSKA Witold JACHYMCZYK Celina JANION Andrzej JERZMANOWSKI (Warsaw University/IBB PAS) Maria M. JEZEWSKA Tadeusz KLOPOTOWSKI Tadeusz KULIKOWSKI Jaroslaw KUSMIEREK I Zofia LASSOTA | Grazyna MUSZYNSKA Grazyna PALAMARCZYK Andrzej PASZEWSKI

11 Irena PIETRZYKOWSKA Joanna RYTKA Piotr P. STÇPIEN (Warsaw University/IBB PAS) Jan W. SZARKOWSKI Lidia D. WASILEWSKA-DAJBROWSKA Bernard WIELGAT Wlodzimierz ZAGÓRSKI-OSTOJA Zona ZAREJBSKA Jerzy ZUK

Representatives of Junior Scientists:

Jacek BARDO WSKI Alicja BE_BENEK Iwona FIJALKOWSKA Jacek HENNIG Ewa KULA-SWIEZEWSKA Piotr MIECZKOWSKI Renata N ATORFF Ewa SLEDZIEWSKA-GÓJSKA

12 Members of Trustee Council of IBB-PAS (a group of former Staff Members of the Institute now residing abroad)

Jerzy Barankiewicz Ryszard Kole Cypros Pharmaceutical Corporation Lineberger, Comprehensive Cancer 2714 Loker Avenue West Center, School of Medicine Carlsbad, CA 92008, USA The University of North Carolina at tel: (619) 929-9500 Chapel Hill, Campus Box 7295 fax:(619)929 80 38 Chapel Hill, NC 27599-7295, USA E-mail: [email protected] tel: (919) 966 11 43 fax:(919)966 30 15 Jadwiga Chroboczek E-mail: [email protected] Institut de Biologie Structural 41, Avenue des Martyrs Magda Konarska 38027 Grenoble Cedex 1, France The Rockefeller University tel: (033 4) 76 88 95 80 1230 York Avenue fax: (033 4) 76 88 54 94 New York 10021-6399, USA E-mail: [email protected] tel: (212) 327 84 32 fax:(212)327 83 70 Witold Filipowicz E-mail: Fredrich-Miescher Institut [email protected] 4002 Basel, Schwitzerland tel: (41 61)69 71 111 Piotr Lassota fax: (41 61)69 73 976 American Cyanamid Company E-mail: [email protected] Lederle Laboratories Division Oncology and Immunology Research Leszek Kleczkowski Section, Department of Plant Physiology 401 N. Middletown Road Umea University, Umea, Sweden Pearl River, NY 10965, USA tel: (46) 90 19 34 39 tel: (914) 732 21 57 fax: (46) 90 16 66 76 fax:(914)732 56 95 E-mail: [email protected] E-mail: [email protected]

Jerzy Paszkowski Wlodzimierz Mandecki Fredrich-Miescher Institut Corporate Molecular Biology P.O. Box 2543 Department 93D, Building 9A CH 4002 Basel, Switzerland Abbott Park, Illinois 60064, USA tel: (41 61)69 79 144 tel: (708) 937 22 36 fax: (41 61)69 73 976 fax:(708)938 60 46

13 Norman Pieniqzek Wlodzimierz Szer Division of Parasitic Diseases New York University Medical School Biology and Diagnostic Branch Department of Biochemistry 4770 Buford Highway NE, 550 1st. Ave. MailstopF 13 NY 10016, USA Atlanta, GA 30341-3724, USA tel: (212) 263 53 20 tel: (404) 488 40 73 fax:(212)263 81 66 fax: (404)488 41 08 E-mails: Andrzej Sledziewski [email protected] B iopharmaceuticals-NovoN ordisk [email protected] 1201 Eastlake Avenue East Seattle, Washington 98102, USA Anna Radomiriska tek (206) 442 67 10 Division of Gastroenterology fax: (206) 442 66 08 University of Arkansas for Medical E-mail: [email protected] Sciences, 4301 WestMarkham Mail Slot 567-1 Ludwika Zimniak Little Rock, Arkansas 72205-7199, USA University of Arkansas for Medical tel: (501) 68 65 414 Sciences, Division of Nephrology fax:(501)68 66 248 4301 WestMarkham E-mail: Mail Slot 501 [email protected] Little Rock, Arkansas 72205, USA tel: (501) 661 1202 ext. 2972 Andrzej Stasiak fax:(501)6712510 Universite de Laussanne E-mail: [email protected] Laboratoire d'Analyse Ultrastructurale Batiment de Biologie, niveau 1 CH-I0I5 Lausanne-Doringt, Switzerland tel: (41 21) 692 24 71 fax: (41 21)692 25 40

14 SCHOOL OF MOLECULAR BIOLOGY - PH. D. STUDIES Head: professor Grazyna Palamarczyk

The Warsaw School of Molecular Biology was initially organized in September 1994 to facilitate attainment of the general requirements for the Ph.D. degree in biological sciences. The board of the School consists of professors: G. Palamarczyk, A. Jerzmanowski and J. Rytka. In 1996-97, 12 students involved in the program have fulfilled the requirements for, and obtained the Ph.D. degree. At present, attendance includes 74 candidates for the Ph.D. degree. The teaching program offers lectures and seminars on various topics. In the last two years lectures on ,,Fungal Genetics and Molecular Biology" and ,,Plant Biotechnology" were organized. In 1996 the School initiated series of seminars on topics related to the major interests of research groups from the Institute. The seminars are supervised by the more experienced and advanced Ph.D. students. Until now seminars concerning: (i) ,,Genome Sequencing", (ii) ,,Transcription" and (iii) Molecular Basis of Plant-Pathogen Interaction" have been completed. Students from the 1-st and 2-nd year of the Ph.D. program attended the Tempus International Course on Diversity and Unity in Biological Sciences which was co-organized in May 1997 by the Warsaw University and our Institute. Fifteen students took part in the International Conference on Signal Transduction in Plants which was held at our Institute in November 1997. All students had several opportunities to present their research achievements. In the last two years 53 papers of which the Ph.D. students were co-authors were published. Majority (47) of the students presented their research at the Third Survey of Research of the Institute (November 1996). Ten students were selected to present their work at the Conference on Molecular Analysis of Microbial Genomes organized in December 1996 by the IBB PAS and the Polish-French Centre. All students attending the Tempus Course (May 1997) presented their research achievements in the accompanying poster sesion. Nine Ph.D. students attended the International Student Conference organized in May 1997 by the University of Gdansk. One of them (Tomasz Bykowski) was awarded by the invitation to present his work both orally and as a poster at the International Student Conference in Berlin. Eleven Ph.D. students took part in the International Conference on Molecular Biology of Plants Under Environmental Stress which was held in Poznan (September 1997). Many students attended individually several local or international conferences related to the topics of their research interests. In each instance presentation of their own results was a prerequisite for participation at those meetings. From its own budget the School offers mini-grants (appr. 3.000 PLN each) to support best research projects presented to the Board by the Ph.D. students. Thirteen applicants in 1996 and twelve in 1997 were awarded.

15 POLISH - FRENCH CENTER OF PLANT BIOTECHNOLOGY Director: professor Stanislaw Lewak

The Polish - French Center of Plant Biotechnology was founded in 1993 by an international agreement between CNRS (French National Center of Science Research), KBN (Polish State Committee for Scientific Research) and PAS (Polish Academy of Sciences) as a continuation of the twin association between IBB and CGM CNRS (CNRS Molecular Genetics Center). The Center is financially supported by the governments of both countries. The funds are destinated for the promotion of joint research programmes on plant molecular biology and genetics. The principal Polish participants are the Institute of Biochemistry and Biophysics of Warsaw, the Institute of Bioorganic Chemistry of Poznari and Warsaw University. Polish research teams cooperate with several CNRS and INRA laboratories and French universities. The supported activities concern the followings areas: 1. Higher plants: plants - microorganisms interactions, regulation of gene expression, nucleic acid metabolism, metabolic regulations. 2. Lower plants (yeasts and filamentous fungi): sequencing and functional analysis of the yeast genome, studies on nuclear - mitochondrial relationships, regulation of metabolic pathways. 3. Plant viruses - studies on transgenic resistant plants. The Center's financial contribution to the Polish - French cooperation consists in supporting short-term (1-3 months) stays (14 persons in 1996, 11 in 1997) and joint research programs (22 grants in 1996, 17 in 1997). Moreover, the organization of six conferences and seminars was partially financed by the Center in 1996-97: (i) French - Polish Conference "Gene structures and expression in plant pathogenes", 18-20 January 1996; (ii) French - Polish Workshop on Bioinformatics, Warsaw, 3-5 June 1996; (iii) French - Polish Seminar "Improving seed quality", Skierniewice, 16-19 July 1996; (iv) French - Polish Seminar "Molecular analyses of genomes of microorganisms", Warsaw, 9-10 December 1996; (v) International Conference "Mechanisms of DNA repair and mutagenesis" on the 100th Anniversary of the discovery of Polonium and Radium, Warsaw, 8-11 October 1997; (vi) French - Polish Seminar "Molecular basis of signal transduction in plants", Warsaw, 28-29 November 1997.

16 INTERNATIONAL COLLABORATION BASED ON FORMAL AGREEMENTS BETWEEN IBB PAS AND FOREIGN INSTITUTIONS

1. The Max-Planck Institute of Molecular Genetics, Berlin (Germany)

2. Department of Medical and Physiological Chemistry, Uppsala University, Uppsala (Sweden)

3. Research Center of Pharmaceutical Chemistry of the Biological Active Compounds of CNR (Italy)

4. Institute of Biological Chemistry, University of Verona, Faculty of Medicine, Verona (Italy)

5. Laboratory of Terpenoid Chemistry, Institute of Chemistry Moldovian Academy of Sciences (Moldova)

6. State Scientific Center Russia - NPO ,,VECTOR", Koltsovo, Novosibirsk Region (Russia)

7. Laboratory of Carbohydrate Chemistry, Zelinsky Institute of Organic Chemistry, Moskow (Russia)

8. Polish-French Center for Plant Biotechnology, located at IBB PAS, by common agreement between PAS, KBN and French CNRS

17 LIBRARY FACILITIES Head: Teresa Zylka, M. Sc.

The Institute Library, established in 1954, has grown to become one of the best in Poland in the field of molecular biology, biochemistry, and related areas. At present it has almost 7,130 books and 10,460 volumes of periodicals. The journal section includes 325 titles, 167 of which are received on a regular basis, and 25 titles by exchange with domestic and foreign institutions. This collection comprises the most important international publications in biology, molecular biology, biochemistry, chemistry, biophysics, genetics, biotechnology, virology. The Library also has a collection of 411 theses (M. Sc., Ph. D., Dr. habilitatus) delivered at the Institute. The Library prepares and distributes annually a list of publications of the Institute. Computer search services include Life Sciences Collection on CD-ROM, Current Contents and selected periodicals in computerized form, as well as access to Internet. Library E-mail address: [email protected]

18 RESEARCH UNITS OF THE INSTITUTE

NEXT PAGE(S) left BLANK DEPARTMENT OF BIOPHYSICS Head: professor Kazimierz L. Wierzchowski

The Department's main research interests concern structural, conformational, kinetic and thermodynamic determinants of intra- and intermolecular interactions in polypeptide and polynucleotide systems, underlying folding in native forms and biological functions of proteins and nucleic acids. Both experimental (optical- and NMR spectroscopy, steady-state and fast kinetic methods, electrophoretic and chromatographic techniques, biochemical assays) and theoretical computer modeling (molecular mechanics and dynamics and quantum mechanical calculations, threading modeling methods) approaches are employed in research projects on selected systems. These projects include: (i) mechanisms involved in binding of Ca2+ by specific protein carriers, (ii) specific interactions responsible for stabilization of a-helical conformation of polypeptides, (iii) early steps and conformational properties of intermediates involved in folding of a trypsin inhibitor (BPTI) , (iv) long range electron transfer between radical redox centers in model peptides and proteins, (v) modeling of protein crystallization and structure of proteins, (vi) molecular interactions in formation of transcriptional complexes in procaryotic systems, (vii) application of recombinant DNA techniques in some of these projects, and (viii) electric field effects on living cells underlying cell electroporation and electrofusion.

1. CONFORMATIONAL PROPERTIES OF CALCIUM BINDING PROTEINS AND THEIR FRAGMENTS

1.1. Structural studies of S100A1 protein

A. Bierzynski, A. Ejchart, G. Goch, H. Kozlowska, E. Soszynska, I. Zukow

Preliminary ID and 2D H'NIVIR spectra of satisfactory resolution have been obtained for S100A1 protein in water, pH 6.2. First samples of N15 labeled protein have been produced in E. coli. Two mutants: Glu32^Gln and Glu73-^Gln of the a subunit of S100A1 protein (built of two a subunits) have been produced in a high expression system of E. coli. In these mutants the first and the second calcium binding loop, respectively, are inactive. Binding constants of La3+ ions to the respective S100A1 mutants in their reduced as well as oxidized forms have been determined by fluorescence measurements. This work is aimed at elucidation of structural transitions of the protein induced by metal binding to each of the loops and at explanation of cooperativity mechanism of metal coordination. Publications: 3214, 3215, 982/A, 63/N

21 1.2. Investigation of an a-helix nucleation by a calcium-binding loop

A. Bierzynski, H. Kozlowska, K. Pawlowski

It was shown previously by us that in the peptide AcDKDGDGYISAAEAAAQNH2 the metal binding loop D1-E12 saturated with lanthanide ions nucleated the a-helical conformation of the C-terminal segment A13-Q16 with extreme efficiency. Comparative analysis of CD spectra of the peptide measured at various temperatures and trifluoroethanol concentrations show that the a-helix content in the A13-Q16 segment is close to 100% (no less than 95%) at low (1°C) and even room (25°C) temperatures. Only at 50°C it drops to about 85%. Theoretical analysis of these results leads to the conclusion that the helix initiation is not described correctly by the existing helix-coil transition theories. Additional effects must be taken operational that make the helix nucleation much more difficult and its initial propagation much more favorable than the current theories would have it. The peptide provides a unique system for studies of spectral and conformational properties of extremely short a-helices. The theory of CD spectra of such helices has been discussed in the light of our results. Further studies of the peptide structure and its dynamics are planned using NMR methods. To produce the N15 labeled material needed for this purpose the gene coding for the peptide has been cloned in E.coli. Publications: 3184,3243

2. LONG-RANGE ELECTRON TRANSFER BETWEEN RADICAL REDOX CENTERS IN MODEL PEPTIDES AND PROTEINS

K. L Wierzchowski, K. Bobrowski, J. Poznanski, K. Majcher

Kinetics of long-range electron transfer (LRET) accompanying various radical transformations involving aromatic (Trp and Tyr), or aromatic and sulphur (Trp or Tyr and Met) amino acids separated by oligoproline bridges ( -Pron-, n = 0-5) has been studied by pulse radiolysis techniques in collaboration with Dr. J. Holcman of the Ris0 National Laboratory (Denmark). Interpretation of the kinetic data in terms of the Marcus theory of distance dependence of the rate of LRET, taking into the account of conformational preferences and dynamics of the model peptides investigated, allowed (i) to demonstrate the involvement in these systems of two electron-transfer pathways: through the peptide backbone (TB) and through space (TS, only in short-bridged peptides with n = 0-2), and (ii) to evaluate the corresponding descriptors, |31B and p1s, of exponential decay of the electronic wave function (K. Bobrowski et al. J.Phys. Chem. 96, 10036, 1992; J. Poznanski, Ph.D. Dissertation, 1996; 3253, 3254). The value of (3rB = 2.5 nm'1 thus obtained, lower by a factor of ~ 3 than the average one for

presently at the Institute of Chemistry and Nuclear Techniques, Warszawa, ul. Dorodna 16

22 LRET in metallo-proteins, indicates that oligoproline bridges (for n > 2 in al\-trans 31 helical type conformation) form very efficient helical chanels for electron transfer and suggests that cc-helical segments in redox proteins may function similarly. More recently we studied kinetics, thermodynamics and radical yields of pulse radiolytically induced Trp* -> Tyr' radical transformation in hen egg-white lysozyme (HEWL) [3207]. HEWL was chosen as a model for these studies because of its well known structure. The main goal of the study was to identify which of the potential Trp'/Tyr redox pairs, formed by six tryptophan and three tyrosine residues present in HEWL, and which of the associated LRET pathways are actually involved in the observed transformation. For this purpose we investigated also NFKyn62-HEWL, a derivative of HEWL in which Trp62 was selectively oxidized with ozone to N'- formylkynurenine, and HEWL(GlcNAc)3, a complex of HEWL with an oligo- saccharide inhibitor triacetylchitotriose, in which Trp62 and Trp63 are known to be completely shielded from outer reagents by the bound inhibitor. In addition, the parameters of LRET were determined as a function of pH and temperature, which were used to perturb the structure of HEWL in a controled way. Analysis of the experimental data allowed us to conclude that in native HEWL the three most solvent- accessible tryptophans: Trp62, Trp63 and Trp 123 are simultaneously involved in LRET, with Trp62 contributing at least ca. 50% to the radical yield. Moreover, electronic coupling between these three Trp acceptor centers with respective Tyr donor centers should be similar to one another in each case, since the overall rate of LRET could be approximated within the experimental error as a single first-order process. To evaluate which of the three tyrosine residues present in HEWL: Tyr20, Tyr23 and Tyr53, form effective Trp'/Tyr redox couples with one or more of the three Trp residues implicated in LRET, we calculated electronic coupling values between all possible Trp/Tyr redox pairs along the corresponding shortest pathways using the PATHWAYS model and crystallographic coordinates of triclinic HEWL. The largest and and approx. equal values were obtained for the Trp62/Tyr53 and Trp62/Tyr53 pairs, and a lower one for the Trpl23/Tyr23 pair. However, molecular dynamics simulations of time variation of electronic coupling for the latter pair showed that, because Trp 123 and Tyr23 are most strongly coupled along a pathway including three through-space (van der Waals contacts) electron jumps, their electronic coupling may occasionally attain a much higher momentary value than that calculated for the static structure. Thus, simultaneous involvement of all three selected Trp/Tyr couples seems probable. Publications: 3207, 3253, 3254

3. MODELING OF PROTEIN STRUCTURE AND FUNCTION

P. Zielenkiewicz, D. Plochocka, A. Kierzek, S. Gavryushov

Analysis of known protein crystal structures shows that interaction energies between monomers alone are not sufficient to overcome entropy loss due to the fixing of protein monomers in the lattice of a growing crystal. Interactions with several

23 neighbours in the crystal are required for the stabilization of the monomers in the lattice. A microscopic model of nucleation and early growth stages of protein crystals has been elaborated, based on the above observations. Anisotropy of protein molecules is taken into account by assigning free energies of association ( proportional to the buried surface area ) to individual monomer-monomer contacts in the lattice. Random walk lattice simulations based on the above model correctly reproduce the speed of movement of a dislocation on the 110 crystal face if the orientational probability is set to 10"5. At this value of the orientational probability, a tetramer, which is the smallest stable complex of the crystal order, appears in lml of solution in the time scale of microseconds. To perform calculations in the timescale of hours a new algorithm was developed, equivalent to the random walk one. With the use of this algorithm size distribution of microcrystals in the timescale of hours was determined. The predicted number of macroscopic crystals is in agreement with the corresponding value obtained in the arrested nucleation experiments. The model can be applied to aid interpretation of light scattering data for crystallizing protein solutions. The fact that the orientational probability predicted by the model is in agreement with this value obtained for the barnase-barstar complex shows that this work can help in the estimation of important characteristics of the protein aggregation process. A three-dimensional model of glycosylase 1 (Tag A) has been proposed. The glycosylase Tag A is a repair enzyme which specifically removes modified adenine residues from DNA. The amino acid sequence of the enzyme is known, but up to now this protein has not been crystallized. Threading methods predict that the sequence of glycosylase I fits to the known structure of erythrocruorin. Based on this the structure of glycosylase I was modeled to be all alpha-helical, in agreement with the observed CD spectrum. The Bogolyubov-Born-Green-Yvon (BBGY) equations were applied to study ionic and mean electrostatic potential distributions around hen egg white lysozyme molecule. The elaborated algorithm allows to apply the BBGY equations to macromolecules of arbitrary shape and charge distribution. The results of calculations for lysozyme were compared with those obtained by Poisson-Boltzmann calculations. It was found that ion-image charge interactions and interionic correlations (both of which are neglected by the PB equation) have a weak effect on the electrostatic potential near charged groups of the protein. They do, however, significantly change the potential and ionic atmosphere at large separation from the macromolecule. Publications: 3153, 3218, 3246

4. MOLECULAR MECHANISM OF TRANSCRIPTION INTIATION IN PROCARYOTIC SYSTEMS

K. L. Wierzchowski, T. Lozihski, W. J. Smagowicz and I. Kolasa

Molecular interactions involved in the formation of transcriptional binary and tertiary complexes have been subject of our studies for more than a decade. In the last years, we were primarily interested in the effects of DNA bending on promoter structure and activity (Lozinski et al., Nucl. Acids Res. 19, 2947-2953, 1991, Lozinski

24 & Wierzchowski, Ada Biochim. Polon. 43, 265-280, 1996). For this purpose a set of consensus-like E.coli promoters bearing An.Tn bending tracts (n = 4-6) in various functional regions (-35, spacer, and -10) were synthetized and their relative strength in vivo and the gross-structure of the open transcriptional complexes with cognate RNA polymerase (RNAP) were investigated in vitro by the PAGE gel shift method. It was found that the most significant effects on the promoter activity (a 3-4 fold decrease in) and the electrophoretic mobility of transcriptional complexes (lowering of) were exerted by T4[-34...-37] and T5[-34...-38] tracts located in the nontemplate DNA strand and overlapping partially the -35 consensus hexamer TTGACA. These effects were interpreted, in connection with the available 3D low-resolution model of RNAP, as a result of the perturbation by the bending tracts of subtle specific interactions between (i) the -35 recognition hexamer and the hypothetical helix-turn-helix motif of the a70 subunit and (ii) the upstream DNA helix with the a subunit of the enzyme. More recently, we studied the kinetics of formation and dissociation of open transcriptional complexes formed on the bent promoters in vitro, to elucidate better the molecular mechanisms underlying the observed effects of promoter DNA bending. The kinetics of both processes was followed spectrofluorometrically, using a fluorogenic y- ANS-UTP substrate, under conditions of abortive transcription initiation. From the kinetic data for the forward reaction, initiated by the addition of RNAP, the characteristic lag-time, x, at which the reaction enters the steady-state regime was determined, and parameters of the two-step reaction model: the equilibrium constant for closed complex formation, KB, and the rate of its isomerization to the open complex, kf dervied therefrom. Comparative analysis of these parameters indicated that DNA bending sequences induce measurable differentiation of their values, the magnitude of which depends on the number and localization of the An.Tn bending tracts within the promoter. Moreover, the relatve strengths of bent promoters in vitro, expressed as a product KB xkf, varied to a larger extent than those measured in vivo. Quantification of open complexes formed in vivo (by KMnO4 footprinting) on the parent unbent promoter a (on the pDS3 plasmid), in the presence and in the absence of rifampicin, demonstrated that the rate of RNA synthesis is not limited by the rate of open complex formation. This observation explained in part the different hierarchy of the promoters established in vivo and in vitro. The rate constants for dissociation of open complexes, kd, proved practically idependent of the presence of DNA bending tracts. This seems to indicate that once an open complex has formed, the promoter DNA therein adopts the same conformation irrespective of its bendability.

For the complex of RNAP with the parent promoter a the variation of kd with temperature and concentration of MgCl2 has also been investigated to evaluate the number N of Mg2+ ions bound at the final stage of its formation. Following a protocol developed by the group of T. Record, we found that N ~ 3, as reported for APR-RNAP complex by Suh et al. (1992); A,PR is the only promoter studied earlier in this regard. We also developed a method for determination of the dissociation-constant of specifically 2+ bound Mg ions from the open complex, Kd, which relies on KMnO4 footprinting and quantitative evaluation of the growing oxidability of T residues within the transcription buble with increasing concentration of MgCl2. Similar values of Kd were found for a

25 number of promoters from the series studied and of the same, mM, order of magnitude as that determined by EPR for T7 RNA polymerase by Woody et al.(1996). In the lattter case Mg2+ ions were shown to be bound to the carboxylic groups of Asp residues located close to the active site of the enzyme. Similarity of Kd's for the two RNA polymerases may correspondingly indicate similar nature of their binding sites for Mg2+. To approach the problem also from the protein side, most recently we overexpressed in E.coli a number of a70 subunit fragments from its 4.2 region bearing a postulated helix-turn-helix domain involved in specific recognition of the -35 promoter hexamer. Publications: in preparation.

5. INHIBITORS OF SERINE PROTEASES FROM CUCURBITACEAE FAMILY

M. Fikus, B. Rempola, J. Topczewska

A gene, coding for a potential chymotrypsin inhibitor, was synthesized, cloned in E.coli and its sequence confirmed. The gene was recloned to a S. cerevisiae secretion - expression vector constructed in our laboratory, pYET. However no inhibitor activity was detected either in the bacterial or in the yeast cloning system. Publications: 3110, 3117, 3145, 3244, 38/N, 40/N, 53/ N, 1089/A

6. ELECTRIC FIELD EFFECTS ON LIVING CELLS

M. Fikus, P. Pawlowski

The mechanical properties of Neurospora crassa cells subjected to a periodic electric field, in the presence of various concentrations of cytochalasin B, were investigated. Shear and extentional deformations were considered and studied separately. Conditions were found under which shear deformations become irreversible. The results were interpreted in terms of rheological models and respective rheological parameters were calculated. The influence of cytochalasin B on the course of extentional deformation of cells was characterized semiquantitatively. Publications: 3109, 3240, 954/A, 1090/A

26 7. STRUCTURE OF THE EARLY FOLDING INTERMEDIATES AND THEIR IMPACT ON THE FOLDING PROCESS.

M. Dadlez, K. Zdanowski, J. Dyczkowski, Z. Podgorska

The main area of interest concerns the conformational properties of early folding intermediates and their impact on the protein folding process. The interest in this largely unexplored area is strengthened by the recently growing recognition that structural deviations in the folding intermediates may underlie many pathological processes in living organisms including humans. Bovine pancreatic trypsin inhibitor (BPTI) is the object of our studies. It is a small protein with three disulfide bonds stabilizing its structure. When these disulfide bonds are reduced the protein unfolds. Starting from the reduced species, re-formation of disulfide bonds can be initiated and the protein refolded. Differently disulfide bonded intermediates can be resolved, quantitated and isolated on a prepatative scale using high pressure liquid chromatography (HPLC). The disulfide rearrangement studies provide a unique insight into the conformational properties of unfolded proteins, not available by classical methods of structure analysis. Following the isolation of a fast folding intermediate 14-38 (i.e. containing the disulfide between cysteines 14 and 38) of BPTI [Dadlez, ML, & Kim, P.S. (1995) Nature Struct. Biol., 2, 674-679] the structure of this intermediate was characterized in a greater detail. It was shown that the structure of the unfolded protein promotes fast formation of this intermediate. By means of site directed mutagenesis a series of BPTI mutants was constructed, expressed in E. coli and studied. It was shown that a group of hydrophobic residues facilitates the search for native fold of the protein. The interactions stabilizing such flexible folding nucleus were characterized thermodynamically. The role of hydrophobic interactions in initiating the folding has been postulated before by theoretical models of the folding process. In collaboration with prof. J. Otlewski from the University of Wroclaw a series of the folding intermediates of BPTI was studied in terms of their affinity to the target enzyme (trypsin). It was shown that specific binding can be detected even for the early folding intermediates including the unfolded protein in folding conditions. It indicates that early in the folding process structural elements are unexpectedly stable. Also in collaboration with the Wroclaw group we have started to employ site- directed mutagenesis and protein libraries selection in studies on the folding and structural properties of proteinases inhibitors and the interactions between proteinases and their inhibitors. Publications: 3170,3190,3237

27 DEPARTMENT OF GENETICS Head: professor A. Paszewski

There are four independent research groups in the Department, all involved in fungal genetics. One group studies the mechanisms of mutagenesis. By using a broad range of cell-division (cdc) and radiation-sensitive (rad) mutations, DNA repair and recombination (in particular the role of DNA polymerases in these processes) are analyzed at the physiological and molecular levels in the yeast Saccharomyces cerevisiae. The second group investigates the regulation of heme biosynthesis and heme- contai-ning enzymes, and the formation and functioning of peroxisomes. The mechanisms of action of regulatory factors such as heme, glucose and oxygen are studied using classical yeast genetics and molecular approaches. Another line of investigation in yeast concentrates on identification of nuclear genes involved in protein transport to mitochondria and their functions in stress response and protein ubiquitination. The regulation of sulphur amino acid metabolism is studied in Aspergillus nidulans and various species of yeasts. These investigations mainly concern the role and regulation of alternative pathways of cysteine and methionine synthesis, in particular the role of the sulphate metabolite repression system. Structural and regulatory genes are cloned and characterized. The interrelations among regulatory circuits governing sulphur metabolism enzymes and folate enzymes are analyzed.

1. GENETIC REGULATION OF SULPHUR AMINO ACID METABOLISM IN FUNGI

A. Paszewski, J. Brzywczy, I. Lewandowska, R. Natorff, M. Piotrowska, M. Siehko, J. Topczewski

Our studies of genetic regulation of sulphur metabolism in Aspergillus nidulans focus on cloning and characterization of structural and regulatory genes. The genes coding for cysteine synthase (cys B), homocysteine synthase (cysD) and cystathionine beta-lyase (metG) were sequenced, their intron/exon structure determined and expression at various conditions of sulphur supply was analyzed. Two other structural genes: cysC and metE, involved in cysteine and methionine synthesis, respectively, are now being characterized. Two sulphur regulatory genes - sconB and sconC - out of four identified, were cloned and analyzed. The former codes for a protein belonging to a large family of regulatory proteins characterized by the presence of several repeats of the"WD" motif. It also contains a newly identified "F-box", encountered in various regulatory proteins, and a nuclear localization signal. The SCONB protein shows a high homology to

28 N.crassa SCON-2 and S.cerevisiae MET30 proteins, both involved in sulphur regulation. The sconC gene encodes a protein of 160 amino acids sharing 51 % identity with the yeast SKP1 protein of central importance to a number of cellular processes, e.g. the cell cycle. The SCONC protein contains the PEST sequence common in rapidly degraded proteins. Its sequence predicts glycosylation and phosphorylation. We have also identified three new genes in A. nidulcms (folA, folB,and folC) which regulate the synthesis of folate-depedent enzymes. Mutations in these genes render the cells intensitive to methionine-mediated repression. A Kluyveromyces lactis gene coding for homocysteine synthase was cloned in S.cerevisiae by complementation of a met25 mutation. It was found that expression of this gene is regulated in S.cerevisiae which indicates that both yeast species have similar systems regulating sulphur metabolism. Publications: 3140, 3147, 3148, 3149, 3232, 3233, 904/A, 1063/A, 1064/A, 1065/A, 41/N, 62/N

2. NUCLEAR GENES INVOLVED IN MITOCHONDRIAL BIOGENESIS IN YEAST

M. Boguta, A. Chacinska, A. Konopinska, K. Pluta, B. Szczesniak

The NAM9 nuclear gene of Saccharomyces cerevisiae was previously reported to code for a mitochondrial ribosomal protein. Using the epitope-tagging technique Nam9p was identified as a protein of 56 kD. The regulation of NAM9 gene expression was studied. The increase in the NAM9 gene dosage was accompanied by the increase in both mRNA and protein. The levels of the NAM9 transcript and protein were both reduced in cells growing on glucose as compared to galactose as a carbon source. Nam9p accumulated at the same level in rho" and rho cells. These results confirm previous data indicating quite diverse regulation of mitochondrial ribosomal protein genes. Cellular extracts were fractionated and, as expected, Nam9p was identified only in mitochondria. When expressed in an in vitro transcription-translation system, Nam9p was imported to isolated yeast mitochondria and no presequence was cleaved. A genetic screen was applied to identify proteins which interact with Nam9p or affect the NAM9 expression. The most interesting one is HsplO4 which promotes the resolubilization of aggregated proteins including the yeast prion-like factor *F (data from the laboratory of S. Lindquist, University of Chicago). The defect of respiratory growth due to the NAM9-1 point mutation Ser82->Leu disappeares when a multicopy plasmid encoding HsplO4 is introduced and does not appear again when the plasmid is lost. A similar effect is observed when guanidine hydrochloride is present in the medium. Inactivation of chromosomal copy of the HSP104 gene prevents the NAM9-1 respiratory defect. Our genetic results suggest that the conformation of Nam9p is dependent on the level of HsplO4 in the cell. Preliminary biochemical experiments show no effect of the HsplO4 level on the cellular localization and amount of the wild type and mutated Nam9p.

29 The GDS1 gene, a suppressor of the NAM9-J mutation, shows genetic interactions with the HSP104 gene, but molecular studies show that it is not involved in HsplO4 expression. We also continue studies of MAF1, the gene of Saccharomyces cerevisiae identified in our lab. The MAF1 is involved in the control of cytoplasmic translation fidelity. The TEF1 gene encoding an elongation factor EFl-a, was cloned as multicopy suppressor of mutation in as well as deletion of the MAF1 gene. MAF1 was tagged by HA epitope at C-terminus and this enabled the identification of Maflp as a protein of 45 kD. Publications: 3094, 3104, 3152, 3193, 3195, 3235, 964/A, 991/A, 1023/A, 1071/A

I 3. MECHANISMS OF DNA REPAIR, RECOMBINATION AND ; MUTAGENESfS IN SACCHAROMYCES CEREVISIAE I "fr I ^ ^- Jachymczyk, J. Zuk, H. Baranowska-Wyszomirska, A. Halas, A. Ciesielski, i Q Z Policinska \ O : O i —I 1. It was confirmed that from the six DNA polymerases discovered in yeast ; cells, only DNA polymerases 8, e and C, are engaged in dark repair of lesions caused by i UV-light and MMS. DNA polymerase 5 is involved in the repair of both types of lesions, while DNA polymerase s and C, only in lesions caused by UV and MMS, respectively. Other polymerases are not involved or play only a minor role in repair. The results obtained are being prepared for publication. 2. Studies on the involvement of the three replicative DNA polymerases in mitotic gene conversion induced by mono- and bifunctional psoralens (and also by UV- light or MMS) revealed that DNA polymerases a and 8 are the main polyrnerases responsible for induced intragenic conversion. DNA polymerase s seems to play minor role in this process. It is possible that DNA polymerase a may also be involved in DNA repair synthesis but only in cases when the opening of new replication forks is necessary for repair. 3. Studies on the influence of mutations in the replicative and nonreplicative DNA polymerases on adaptive mutations in the cells of Saccharomyces cerevisiae were continued. We found that thermosensitive mutation in the POL2 gene encoding DNA polymerase e increased the frequency of adaptive mutation in a similar manner as found earlier for DNA polymerase 8. A similar effect was observed also in strains with deletions in the MSH3 gene responsible for mismatch repair. Mutations in other DNA polymerases, including the essential DNA polymerase a and the inessential DNA polymerases (3 and C, revealed no effect on this process. Analysis of DNA sequences in the revertants showed that in all cases the obtained reversions resulted from a single nucleotide deletion most often in sequences having short homopolymer tracts. The results obtained suggest that errors arising during DNA elongation and their persistence in mutants deficient in mismatch repair activity seem to be the source of the adaptive mutation appearence in studied cells. Publications: 3206, 1016/A, 1072/A

30 4. THE HEME AND HEMOPROTEINS BIOSYNTHESIS IN YEAST Saccharomyces cerevisiae.

J. Rytka, M. Gora, E. Grzybowska, T. Zolqdek

Heme is a molecule used to build the cytochromes and other hemoproteins in cells of practically all organisms. A key enzyme in heme production is ferrochelatase, a mitochondrial inner membrane-bound enzyme, that catalyzes the last step in heme biosynthesis. By site-directed mutagenesis we have constructed single- and double- residue substitutions of 10 evolutionarily invariant amino acids in the yeast ferrochelatase. The comparison of the effects of these mutations on in vivo heme synthesis with the enzyme defects measured in vitro indicates that the three invariant residues: H-235, D-246 and Y-248 serve as ligands for ferrous ion. The glutamyl residue E-314 appears to be critical for catalysis. The wild type and mutant ferrochelatases were overproduced in E. coli. They constituted 7-8% of the total cell protein and were found in the soluble fraction. This system will be used to purify the wild type and mutant enzymes for the elucidation of its three-dimensional structure. To identify the putative membrane-anchoring fragment of yeast ferrochelatase a hybrid gene was constructed encoding the chimeric S. cerevisiae - B. subtilis - 5. cerevisiae ferrochelatase. The gene is expressed in S.cerevisiae and E.coli cells. The localization and properties of the chimeric enzyme are under studies. Mutants of S.cerevisiae accumulating uroporphyrin (UP) and protoporphyrin (PP) were used as a tool for studying the mechanism of phototoxicity of porphyrins. A possible mechanism of UP photosensitizing activity was proposed. The yeast was also used as a model eukaryotic organism to study the uptake and phototoxic effect of diamino acid derivatives of porphyrins used in tumor photodynamic therapy. It was shown that HpD(Arg)2 and PP(Met)2(Arg)2 sensitize yeast cells to photodamage but have no mutagenic effect on nuclear or mitochondrial genomes. Collaboration : prof. R. Labbe-Bois, Institut Jacques Monod, Paris, France and prof. A. Graczyk, Military Technical University, Warsaw. Publications: 3100, 3126, 3129, 3130,3217,3219, 3227

5. THE SEARCH FOR THE FUNCTION OF THE ORF YIL026C

A. Kurlandzka, M. Wysocka, J. Rytka

In our studies on the mechanism of induction of peroxisomes proliferation by oleic acid a search for a putative fatty acid binding protein (FABP) in S.cerevisiae led to the discovery of a new gene named IRR1 from: irregular cell behaviour (ORF YIL026c), coding for an essential protein of unknown function. We investigated physiological effects of changing the transcription level of the gene after having fused IRR1 with a regulatory catalase A gene promoter. These studies let us conclude that a lowered expression of IRR] influences zygote formation, spore germination and colony formation. Preliminary results indicate that Irrlp may participate in cell-cell and/or cell-

31 substratum interactions. The cytoplasmic localization of Irtip (verified by immunofiuorescence microscopy and Western blotting) suggests that this protein may be an element of a cell outside-to-nucleus signal transduction pathway. Further investigations will include the isolation of proteins interacting with Inip. Publications: 993/A, 73/N

6. MECHANISM OF PROTEIN DISTRIBUTION TO VARIOUS CELLULAR COMPARTMENTS IN Saccharomyces cerevisiae.

T. Zolqdek, B. Gajewska, J. Kaminska.

Two forms of Mod5p, a tRNA modification enzyme, are found in three cellular compartments: mitochondria, cytoplasm and nucleus. Mod5p-I, found in mitochondria and cytoplasm, is responsible for isopentenylation of A 37 in mitochondrial and cytoplasmic tRNAs. Alteration of the subcellular distribution of Mod5p-I affects tRNA mediated suppression. Altered suppression efficiency was used to select mdp mutants which decrease the mitochondrial pool of Mod5p-I. The gene complementing ts phenotype of mdpl mutant was found to be identical to RSP5 encoding ubiquitin- protein ligase. The four mdpl mutations characterized by sequencing are located in RSP5 sequence encoding hect domain that is highly conserved in the family of ubiquitin-protein ligases. The ts phenotype of four mdpl mutants is suppressed by pmal mutations located in the gene encoding plasma membrane H-ATPase. Further characterization of mdpl revealed that these mutants are pFl sensitive, hypersensitive to hygromycin B, cycloheximide, paromomycin and osmotic stress. These phenotypes are not suppressed by pmal. The pmal alleles alone cause pH sensitivity of cells and resistance to cycloheximide. The results indicate that the Rsp5 ubiquitin-protein ligase in addition to its involvement in mitochondrial/cytosolic protein distribution may influence the functioning of plasma membrane H+-ATPase and other plasma membrane transporters. Collaboration: prof. A.K. Hopper, Hershey Medical Center, The Pennsylvania State University, Hershey, PA, USA Publications: 3193, 3195, 3211, 986/A, 989/A, 990/A, 1024/A, 1030/A, 45/N DEPARTMENT OF LIPID BIOCHEMISTRY Head: professor Tadeusz Chojnacki

The continuing interest of this Department in studying the biosynthesis and function of polyisoprenoids is represented by two groups covering differrent aspects of this problem. One directed by professor Grazyna Palamarczyk, is studying the regulation of formation of polyprenols, dolichols and steroids in yeast, and dolichol-dependent transglycosylations in filamentous fungi. The team succeeded in constructing stable transformants of Trichoderma resei with elevated protein secretion caused by the increased number of copies of dolichol mannose phosphate synthase. Other achievements concern the role of farnesyl diphosphate synthase in the biosynthesis of dolichols in yeast. A mechanism of chain termination dependent on the interaction of farnesyl diphosphate synthase with endoplasmic reticulum membranes was postulated. In the other group directed by professor Chojnacki the studies on the characterization of protein prenyl transferases (farnesyl protein transferase and geranylgeranyl protein transferase II) are being continued. These studies revealed some differences in the properties of enzymes of plant and animal origin. It was demonstrated that dithiothreitol can serve as an artificial acceptor in the enzymic reaction of prenylation. In the studies on the occurrence of polyprenols and dolichols leaves of several species of plants belonging to the families Apocynaceae and Verbenaceae were examined as well as tissues of other plants, and liver from a number of bat species. The studies on the biosynthesis of ubiquinone and plastoquinone are also being continued.

1. OCCURRENCE AND FUNCTION OF TERPENOID LIPIDS IN PLANT TISSUES: STUDIES ON STRUCTURE AND BIOSYNTHESIS

71 Chojnacki,,/. Hertel, W. Jankowski, B. Kazimierczak, M. Szymanska, E. Swiezewska, M. Wanke, Vo Si Hung

Studies on the prenylation of proteins in plants were continued. The properties of the Rab prenyl transferase from plants were studied using the Rab3a protein as the acceptor. The trials aimed at the purification of the prenyltransferase are carried on. A new stage of investigation of the biosynthesis of plastoquinone and ubiquinone in spinach cells was started, focusing on the selective transport of these lipids following their biosynthesis in endoplasmic reticulum-Golgi system: PQ towards chloroplasts and UQ to mitochondria. Differences in the transport mechanisms of these compounds were postulated. Continuing our studies on the occurrence of polyprenols in plants through survey of polyprenol patterns was conducted on a large number of Firms mugo plants belonging to a population established 20 years ago in the Powsin Botanical Garden, PAS. It was

33 shown that the polyprenol pattern is unchanged despite great anatomical differences among the plants. A through examination of a large group of fruit trees belonging to Rosaceae (mainly Cornus, Mains and Pnmus) confirmed the previous finding on the constant type of polyprenol pattern in leaves of these plants: polyprenols composed of 19-20 isoprene units are dominating and in genus Pnmus prenologues composed of 35-40 isoprene units are also present. In leaves of a large number of species belonging to the Combretaceae family the presence of polyprenols composed of 20-60 isoprene units was confirmed. In the majority of the species the long chain polyprenols were in the form of acyl esters; only in C. molle they occurred as free alcohols. In leaves of this plant their content was ca. 4% of dry weight. Polyprenols were found in Coluria geoides root tissue culture (cooperation with dr. Olga Olszowska, Warsaw Medical Academy). Studies on the structure of these polyprenols and on the dynamics of their accumulation are in progress. Preliminary search for polyprenols in mosses was performed with negative results; in lichens only trace amounts were found in some species. On performing a similar search in xylem of various trees only hepta-, octa- and nonaprenols were found exclusively in Betulaceae (cooperation with the PAS Institute of Dendrology, Kornik). Leaves of a large collection of Apocynaceae and Verbenaceae were studied in order to extend the previous finding of the presence of large amount of very long chain polypenols (ca. 100 isoprene units) in African samples from Ghana of Plumeria alba (Apocynaceae) and Tectona grandis {Verbenaceae). Very long chain polyprenols were not found in any of about 100 other species of these families growing in Hawaii (cooperation with the Honolulu and the Waimea Botanical Gardens in Hawaii). Livers of various species of bats were studied for the presence of polyprenols and dolichols considering them as a possible indicator of enviromental pollution and an indicator of ageing. A new type of natural dolichol mixture was observed in this material. Trials to achieve low-temperature cyclization of polyprenols were continued in cooperation with the team of the Institute of Organic Chemistry, Academy of Science of Moldova, Kishiniev. Publications: 3122, 3127, 3157, 3176, 3177, 3183, 3242, 928/A, 938/A, 939/A, 940/A, 947/A, 985/A, 986/A, 1000/A, 1031/A, 1082/A, 37/N, 65/N, +3238, +3239

2. POLYPRENOL FORMATION IN THE YEAST SACCHAROMYCES CEREVJSIAE: EFFECT OF FARNESYL DIPHOSPHATE OVEREXPRESSION

A. Szkopinska, K. Grabinska, D. Grabowska, G. Palamarczyk

We demonstrated that cell extracts and membrane fraction from Saccharomyces cerevisiae catalyze in vitro biosynthesis of dehydrodolichols (a- unsaturated polyprenols) whereas in vivo, yeast synthesize dolichols (a-saturated polyprenols). We have also presented evidence that at least in S. cerevisiae the main pool of dolichylphosphate originates from the phosphorylation of dolichol and that this

34 process must be preceded by a-saturation. Investigating the role of farnesyl diphosphate (FPP) synthase in polyprenol biosynthesis we found that FPP is the allylic ,,starter" for the biosynthesis of dolichols in yeast. Based on the results of overexpression in the mutant perturbed in FPP-and squalene synthase activity of the FPP-synthase encoding gene and overexpression of a DNA fragment encoding a ca. 130 aa fragment containing the first aspartate rich domain of FPP synthase, we postulate that in the yeast enzyme the motif for polyprenol chain termination resides within, or in the vicinity of, the first aspartate rich, conserved domain. An interaction between two monomers of FPP- synthase (by two hybrid system) and co-immunoprecipitation of the synthase, which is a cytosolic enzyme, with yet unidentified membrane protein were demonstrated. Concomitantly the work on the involvement of peroxysomal structures in biosynthesis of polyprenols was carried on.

3. CONSTRUCTION OF TRICHODERMA REESEI STRAINS EXHIBITING ELEVATED PROTEIN SECRETION

J. Kruszewska, U. P erlihska-Lenart, A. Janik, G. Palamarczyk

We have demonstrated earlier that integration of a yeast S. cerevisiae DNA fragment containing the DPMI gene (encoding mannosylphosphodolichol (MPD)- synthase) under its own promoter, into the Trichoderma reesei genome results in elevated secretion of cellulytic enzymes by this fungus. More recently we established that a similar effect can be reached if the yeast gene is introduced under T.reesei pki promoter and cbh2 terminator. Four stable transformants with tandem integration of various copy numbers of the yeast DPMI gene were isolated. They exhibited moderately elevated MPD-synthase activity and secreted up to seven fold higher amounts of total proteins as compared to the control (the host strain transformed with the vectors bearing no DPMI insert). Northern analysis of CBHI, the major secretory protein, transcript show no differences in the transcription level between the transformed and the host strains. Comparison of the level of glycosylation of the secreted proteins in the transformed and parental strains indicates that integration of the S. cerevisiae DPMI gene into the T. reesei genome does not lead to hyperglycosylation of the secreted proteins, however, fungal system is capable of glycosylating of the increased amount of the protein. Concomitantly the effect of DPMI overexpression was studied in the thermo- sensitive secretory (sec) mutants of Saccharomyces cerevisiae. Thermotolerance was aquired due to the transformation with DPMI gene in two of the six sec mutants tested. Some of them exhibited also 5-6 fold increase in active invertase secretion. Studying the early reactions in T.reesei protein glycosylation we cloned cDNA coding for guanyltransferase, the enzyme responsible for the last reaction in GDPMan synthesis, as a supressor of the dpml gene mutation. Publications: 3132, 3134, 3217, 929/A, 930/A, 934/A, 935/A, 1060/A, 1061/A, 1062/A

35 DEPARTMENT OF M1CROBIAL BIOCHEMISTRY Head: professor Maria Danuta Hulanicka

There are three independent research groups in the Department, all involved in studies of the mechanisms of gene expression. One line of research conducted by prof. Hulanicka s group concerns regulation of genes involved in sulphur metabolism in enteric bacteria. Studies on the new regulatory gene, cbl, revealed the essential role of the gene product for assimilation of sulphonate sulphur. A new mechanism engaging concurrent action of two transcriptional regulators, CysB and Cbl, in the expression of the tan operon (required for taurine utilization) was shown experimentally in vivo and in vitro. Another line of studies involved isolation of several cysB mutant alleles and characterization of their products by in vitro interactions with responsive cys promoter regions. Studies on genome expression of potato leafroll virus were continued. The full length cDNA copy of viral genome under 35S promoter was constructed and its biological activity was demonstrated. One of the projects realized by dr. Cegtowski's group concerns the functional analysis of stability systems encoded by the plasmid pSM19035 from Gram-positive bacteria. It was found that (i) better-than-random distribution of pSM 19035 into daughter cells is assured by a plasmid encoded addiction system which acts post- segregationally by killing piasmid-free cells, and (ii) plasmid copy number control and better-than-random plasmid segregation are under common regulation mediated by the product of gene co. The other project of this group, supervised by dr. J. Bardowski, is focused on the genetics of sugar metabolism in lactocococci - biotechnologically important bacteria which are widely used in the agro-industry. A new, chromosomally encoded, cellobiose inducible pathway of lactose catabolism was discovered. On the other hand, gene(s) involved in starch degradation were found to be plasmid-encoded. DNA sequencing of genes involved in the utilization of both sugars is under way. The group of prof. T. Klopotowski has been continuing studies on the regulation of expression of cspC - one of the genes of cold shock regulon in Escherichia coli K12. It is involved in transcriptional control of the wca operon encoding for enzymes of exopolysaccharide synthesis. Four transcriptional start sites for cspC were identified. All of them precede a 47 amino acid open reading frame that is located upstream of cspC and may also play a role in bacterial response to low temperature. A model was proposed to explain toxic efect of the mutations shortening the mRNA transcribed from the orf-47 cspC promoter in the absence of CspC. An independent study on D-amino acid racemization in E. coli has been completed. A new subject initiated last year in this group by dr. M. Lobocka concerns mechanisms of active partition of low copy number plasmids to daughter cells at cell division. PI plasmid prophage of £ coli serves as a model system in these studies. Partition of PI depends on two plasmid genes par A and parB, and a centromeric site parS to which ParB protein binds. The main goal of research was to separate

36 autoregulatory functions of partition proteins from their partition functions and to identify host factors essential for PI partitioning. Mutational analysis of par A led to a conclusion that interactions of plasmid partition proteins involved in autoregulation are not essential for partitioning. A method was developed to isolate host mutants defective in PI plasmid partition. Analysis of mutants obtained is in progress to identify host proteins involved.

1. FUNCTIONAL ANALYSIS OF PLASMID pSM19035 FROM GRAM- POSITIVE BACTERIA

P. Ceglowski, I. Kern, I. Sitkiewicz, U. Zielenkiewicz

The low-copy-number and broad-host-range plasmid pSM19035 and its derivative pDB 101 are 1000-times more stable in Gram-positive bacteria of low G+C content than expected for random segregation. Two regions (SegA and SegB) involved in the stable maintenance of pDBlOl have been identified previously. The SegA region encodes a site-specific recombinase which, by resolving plasmid oligomers into monomers, maximizes plasmid random segregation. The SegB region assures better- than-random plasmid segregation. It consists of four genes: 8, e, C, and co. The mode by which this region acts remained, however, obscure. The aim of our present work is to determine how genes from the SegB region act and how they are regulated. The product of gene 8 contains motifs present in a family of ATPases involved in active plasmid partitioning, however, the major stabilization function of the SegB region is dependent on the activity of gene products s and C,. Using various genetic and biochemical methods we obtained results suggesting that genes s and C, encode a plasmid addiction system which acts by post-segregational killing of plasmid-free cells. Similar systems described for other plasmids consist of a stable toxin and an unstable antidote. An inherent property of these systems is that the antidotes form complexes with the toxins, thereby neutralizing the latter in plasmid- carrying cells. Using the yeast two-hybrid system we obtained evidence that the products of genes s and C, interact directly at the protein level. Gene regulation within the SegB region was studied in B. subtilis with the use of transcriptional fusions of the putative promoter regions and a promoterless p- galactosidase gene. Transcription of the gene 8 and the putative operon co-s-^ appeared to be repressed by plasmids carrying only gene co. DNA regions preceding genes 8 and co contain a set of 7 bp direct repeats which might be a target of the gene product co. Similar repeats are present in the 5' region of the copy control gene copS. The promoter of copS gene also was found to be repressed by plasmids containing gene co. We conclude from these observations that both the plasmid copy number control and the better-than-random segregation of pSM19035 are under common regulation mediated by the product of gene co. Publications: 35/N, 61/N, 69/N, 970/A, 1043/A

37 2. GENETICS OF LACTOSE AND STARCH CATABOLISM IN LACTOCOCCUS

J. Bardowski, T. Aleksandrzak, K. Lewicka and P. Cegtowski

Lactococcus belongs to lactic acid bacteria which are of high biotechnological importance since they are widely used in the agro-industry, particularly in the dairy industry. Plants are believed, however, to constitute a natural niche for strains of this genus. Since (i) lactose is the main sugar in milk and (ii) starch serves as the main storage carbohydrate in plants, we focused our research on the catabolic pathways of both sugars. In lactococci the capacity to use lactose as a carbon source is encoded by plasmid genes. Once the plasmid is lost the strain hardly grows on a medium supplemented only with lactose. Using a plasmid-free strain we found, however, that cellobiose induces a new pathway of lactose catabolism. To identify genes involved in this pathway we applied the method of random mutagenesis with the pGhost9::ISSl insertion plasmid which integrates randomly into bacterial host DNA. Through Southern blot analysis the transformants with disrupted genes involved in lactose metabolism were divided into several distinct groups. From each group of transformants the chromosomal DNA regions flanking the integrated plasmid were sequenced. In two of them the deduced amino acid sequences appeared to be homologous to CcpA (catabolite control protein) from Bacillus subtilis. CcpA is responsible for a global regulation of genes involved in various metabolic processes. This indicates that similar global control mechanism(s) might be also operative in Lactococcus. Regarding the starch degrading activity of lactococci, the biochemical and genetic characterization of the amylolytic enzyme produced by L. lactis IBB500 revealed that: (i) the enzyme has a broad pH- and temperature optima and is accumulated at the early stationary phase of growth, (ii) starch induces while glucose represses its production. Analysis of the plasmid content in L. lactis IBB500 showed that several plasmids exist in this strain and the presence of one of them is correlated with the ability of the strain to degrade starch. Through deletion-subcloning analysis the a my gene was localized within a 1.5 kb DNA fragment. A partial DNA sequence of this gene reveals that its putative protein product exhibits significant homology to an isoamylase from Pseudomonas amyloderamosa. Publications: 3223, 77/N, 78/N, 891/A, 893/A, 894/A, 895/A, 986/A, 898/A, 899/A, 900/A, 901/A, 902/A, 980/A, 981/A, 984/A, 998/A

3. REGULATION OF SULPHATE METABOLISM IN Enterobacteriaceae

D. Hulanicka, M. Hryniewicz, R. Iwanicka-Nowicka, A. bochowska, T. Bykowski

Inorganic sulphur is assimilated by enteric bacteria predominantly via cysteine biosynthetic pathway, involving over 20 genes and controlled by the CysB transcriptional regulator. Recently, we have identified and characterized the cbl gene,

38 encoding a new regulatory protein engaged in sulphur metabolism of E. coli. We established that the functional Cbl protein is required for assimilation of organic sulphonates as a sulphur source. In particular, expression of the tauABCD operon encoding genes involved in the utilization of taurine was shown to depend on both the cysB and cbl gene functions. We performed studies in vitro on the interaction of CysB and Cbl with DNA of the tau promoter region. Studies on the in vivo expression of the tauA-lacZ and tauD-lacZ fusions confirmed that both CysB and Cbl were required for the tau operon expression. This concurrent action of two transcriptional regulators in activation of the tau promoter represents a new paradigm in the regulation of sulfur metabolism-related genes and will be a subject of further investigation. Another line of research is aimed at the elucidation of interaction of CysB protein with RNA polymerase (RNAP). We have isolated several suppressors of rpoA341 mutation (K271E substitution in RNAP a subunit, abolishing transcription initiation at cysP promoter) as Cys+ clones. The major class of these clones comprised pseudorevertants ofrpoA341 allele containing an identical E271A change in the RNAP a subunit. This finding suggests a crucial role of the a subunit residue 271 for CysB- dependent activation of the cysP promoter (possible contact site for CysB). We also isolated several cysB mutant alleles and selected those encoding CysB protein capable of binding to DNA but defective in transcriptional activation of cys promoters (so called cysB positive control mutants, cysBpc). Proteins encoded by three cysBpc alleles were overexpressed and partially purified for in vitro studies on DNA binding mode, cofactor(s) effects and ability to form tertiary complexes with RNAP. These studies are currently under way. Collaboration: Mikrobiologisches Institut, Eidg.Techn. Hochschule ETH-Zentrum, Zurich, Switzerland Publications: 3173, 3194, 3260, 46/N, 66/N, 937/A, 1075/A, 1081/A

4. REGULATION OF POTATO LEAFROLL VIRUS GENOME EXPRESSION

D. M. Hulanicka, E. Sadowy, M. Juszczuk

Potato leafroll virus (PLRV) causes significant potato crop losses world-wide. Our studies on this virus can be divided into the following sub-projects: 1. The construction of infectious clones is the best tool for studies of the molecular biology of RNA viruses. The full length cDNA copy of the viral genome was cloned under transcriptional control of the cauliflower mosaic virus 35S promoter and terminator sequences in a binary vector pBinl9 (plasmid pBPF). Eleven non-viral nucleotides present between the transcription start and the beginning of the genome were removed by site directed mutagenesis (plasmid pBOK). Both plasmids were introduced into Agrobacteriunt tumefaciens LBA4404. The viral coat protein was present in the leaf discs inoculated with agrobacteria containing pBPF or pBOK and was absent in leaf discs inoculated with LBA4404 as shown by Western analysis. This result confirms the ability of the 35S dependent transcript synthesied in planta to

39 initiate a replicative cycle of the virus. In parallel, agroinoculation of whole plants with the same strains of agrobacteria was performed. No signal from the viral coat protein could be detected. 2. PLRV uses several strategies to translate its genome. In the recent studies we have analysed the role of the leader sequence (LS) of PLRV genomic RNA (gRNA) on the expression of ORFO and ORF1, P28 and P70 proteins, respectively. Two sets of plasmids were constructed in order to evaluate the role of in an LS in vitro system. The results indicate: a) LS of gRNA decreases the expression of both genes; b) LS affects the ratio of protein 28K to 70K; c) cap has not effect on the level of protein expression. The other line of our studies concerns a putative protease activity of P70. The characteristic motif for serine protease has been found in this protein. However, the processing of P70 has not been demonstrated before. In order to check the protease activity of this protein two sets of plasmids carrying viral cDNA of ORF1 under T7 promoter were constructed. In one set the putative active centre of serine protease was mutated. The results indicate that the protein P70 is autoprocessed. The autoprocessing of P70 was confirmed in experiments with plasmids carrying the full length PLRV genome. Collaboration: dr B. Gronenborn, Jnstitut des Sciences Vegetales CNRS, Gif-sur- Yvette, France, dr J. van Heuvel Institute for Plant Protection, IPO-DLO, Wageningen, The Netherlands. Publications: 3144, 3196, 42/N, 50/N, 948/A, 949/A, 953/A, 955/A, 956/A, 957/A, 1066/A, 1067/A, 1068/A, 1069/A, 1075/A

5. MECHANISMS OF ENTEROBACTERIAL ADAPTATION TO ENVIRONMENTAL CHANGES

T. Klopotowski and K. Krajewska-Grynkiewicz

We have been using expression of the wca operon (formerly cps) in the study of low temperature effects on the bacterium Escherichia colt. We discovered that the cspC gene product (belonging to the cold shock family of proteins with the evolutionarily conserved Y box motif) exerts a positive effect on wca expression. During the past two years we concentrated on transcription of the cspC gene. We identified four transcription initiation sites all preceding a 47 amino acid open reading frame located upstream of the cspC gene. We suppose that the protein product of the postulated orf-47 gene may play a role in bacterial response to low growth temperature. We found that mutations shortening the mRNA started by orf-47 cspC promoter affect cell viability in the absence of CspC protein. Similar effect was described by other authors for mutations shortening the cspA mRNA. Both effects may result from a requirement for specific RNA chaperone activity to prevent incomplete mRNA toxic effect. The CspA protein provides such an activity for the truncated cspA mRNA as already documented. In our case we suspect that CspC protein would act as such chaperone preventing loss of cell viability in cells carrying plasmids with truncated orf-

40 47 cspC mRNA. We presume that the damage caused by low temperature resides on increased stability of internal double helical segments of RNA molecules. We finished our study on D-amino acid racemization in E. coli with a report on the control of D-amino acid dehydrogenase gene dadA by LRP pleiotropic regulatory protein. Publications: 3102,1087/A 1091/A

6. MECHANISMS OF ACTIVE SEGREGATION OF LOW COPY NUMBER PI PLASMID TO DAUGHTER CELLS AT CELL DIVISION

M. Lobocka, A. Jcmowicz

I. Partition of PI plasmid to daughter cells at cell division depends on plasmid-encoded partition proteins, ParA and ParB, and a as-acting centromere analog, parS. Plasmid partition proteins are postulated to attach a plasmid molecule to as yet unidentified chromosome partition machinery. Binding of ParB to parS is an initial step of partitioning. Excess ParB or changed context of parS may lead to a partition dysfunction - destabilization of certain parS plasmids by ParB. Destabilization is caused by transcriptional silencing by ParB of genes flanking the centromere. Partition defects in destabilization deficient parB mutants suggest that the transcriptional silencing is a step of partitioning. To identify host proteins involved in PI partition we used the phenotype of destabilization to develop a method for selection of chromosomal mutants defective in this partition dysfunction. Some of the mutants obtained influenced stability of wild-type mini-Pi, as expected. A partition marker - mini-P7 plasmid was constructed to accelerate screening of mutants for defects in partitioning. It carries the /aclq gene, the replication and partition region of P7 plasmid related to PI, and the multimer resolution system of PI, lox ere. II. We continued studies on ParA protein. ParA has a weak ATPase activity believed to provide energy during plasmid movement. ParA can also repress the expression of parAB operon by binding to its operator site (parPO). Both activities are stimulated by ParB. To study partition it is crucial to separate regulatory functions of Par proteins from functions directly involved in partitioning. We characterized parA mutants isolated previously by testing the ability of mutant proteins to bind to the parPO in vitro and to repress the expression of parAB operon in vivo. One of the mutants appeared to have properties of superrepressor, independent of ParB in its ability to repress the expression of parPO completely. Our results indicate that the superrepressory properties of mutant ParA and its altered interaction with ParB are caused by increased affinity to ADP. Partition activity of mutant ParA is only slightly altered. This implies that the interaction of ParA with ParB required for the stimulation of ParA binding to parPO is not essential for partitioning. Collaboration: dr. Michael Yarmolinsky, National Cancer Institute, NIH, Bethesda, USA Publications: 1088/A, 1092/A

41 PL0001645 DEPARTMENT OF MOLECULAR BIOLOGY Head: professor Jarostaw Kusmierek (till 31.12.1997 professor emeritus Celina Janion)

The majority of our studies are centered on (i) mechanisms of mutagenesis and DNA repair (including MFD) in Escherichia coli, Ml3, and lambda phages; (ii) inhibitory and miscoding properties of modified bases in DNA; (iii) synthesis and properties of pyrimidine nucleosides and nucleotide analogues vvith potential anti-tumor, anti-virus and anti-parasite activities, including their conformation and substrate/inhibitor properties in some enzyme systems of relevance to chemotherapy; (iv) molecular mechanisms of PUVA (psoralen + UVA) treatment in psoriasis photo-chemotherapy in particular its action on cell membrane; (v) specificity and methods for assays of N-alkyl-purine DNA glycosylase. The spectrum of mutagens tested includes: MMS, DMS, ultraviolet or halogen light and hydroxyl radicals. The enzymes and repair systems investigated include: DNA polymerases and the proofreading activity of DNA pol III, UvrABC-endonuclease, mismatch repair system, and methyl DNA glycosylases. Much attention is focussed on the role of UmuDC proteins in mutagenesis (dependent and independent on DNA replication) and DNA repair, and on the effect of the Tn/0 transposon on the survival and mutation frequency of halogen light irradiated bacteria. A new class of nucleosides containing C(2)-hydroxymethyl-ribose (hamamelose) was synthesized, and it was found that uracil and 5-fluorouracil derivatives show a significant antitumor activity. It was found that 2CDA (2-deoxy-2-chloro-adenosine) an anti-lymphoid drug does not induce mutations, when incorporated into DNA, but significantly inhibits DNA replication. In studies with oxidized Ml 3 DNA it was found that Fapy- (formamidopyrimidine)-residues in DNA selectively inhibits DNA synthesis, and the effect depends on the neighbouring sequences and the DNA polymerase tested. Highly unstable derivatives of lecithin-psoralen adducts were characterized and their role in PUVA photochemotherapy is being studied.

1. MUTAGENESIS AND REPAIR OF DNA PL0001646

C. Janion, E. Grzesiuk, A. Fabisiewicz, B. Tvdek, J. Ciesla, M. Grqziewicz, A. Wqjcik, E. Speina

The discovery that the mfd gene codes for a transcription-coupling repair factor (TRCF) prompted us to re-investigate the MFD (mutation frequency decline) phenomenon in E.coli K-12 strain when mutations were induced by ultraviolet light, halogen light or MMS-treatment. These studies revealed that: (i) the process of MFD involves the

42 proofreading activity of DNA pol III and the mismatch repair system, as well as, TRCF and the UvrABC-excinuclease (ii) a semi-rich plate test may be replaced by a rich liquid medium, (iii) the ToT pyrimidine dimers are the lesions excised with the highest activity, and (iv) overproduction of UmuD(D'C) proteins leads to a great increase in mutant frequency in irradiated and MMS-treated cells. The role of mismatch repair (MR) in MMS-induced mutagenesis is obscured by the fact that the spectra of mutational specificity are different in bacteria proficient and deficient in MR. It has been found that transposons TnlO (and Tn5) when inserted into chromosomal DNA of E. coli influence the phenotype lowering the survival and frequency of mutations induced by UV or halogen light irradiation. This is connected with a deficiency of UmuD(D') and UmuC proteins. Transformation of bacteria with plasmids bearing the nmnD(D')C genes, suppresses the effects of the transposon insertion, a phenomenon which has not been described before. Single-stranded DNA of M13mpl8 phage was oxidized in vitro by a hydroxyl radical generating system including hypoxanthine/xanthine oxidase/Fe3+/EDTA, and it was found that Fapy-Ade, Fapy-Gua, 8-oxyAde and thymine glycol were the main products formed. Replication of the oxidized template by T7 phage DNA polymerase, Klenow fragment of polymerase I, or polymerase beta from bovine thymus has revealed that oxidized pyrimidines are stronger blockers than oxidized purines for T7 phage and Klenow fragment polyinerases and the blocking potency depends on the neighbouring bases and on the type of polymerase. Studies of mutagenic lesions in phage Ml3 DNA after DMS+NaOH treatment strongly indicate that Fapy-7meA is a mutagenic base. The loss of 7meAde, the precursor of Fapy-7meA, from DNA of M13 resulted in a decline in mutagenic potency at a rate proportional to the amount of bases removed. Publications: 3106,3108,3133,3169,3174,3201,3230, 905/A, 906/A, 923/A, 961/A, 962/A, 963/A, 1004/A, 1008/A, 1014/A, 1015/A, 1020/A, 1022/A, 1035/A, 1036/A, 1037/A, 1038/A, 1039/A, 1040/A, 1041/A, 1042/A

2. CHEMICAL BASIS OF MUTAGENESIS AND REPAIR OF CYCLIC BASE ADDUCTS

J. Kusmierek, A. Bukowska-Maciejewska, E. Borys

2.1. Miscoding properties of modified bases.

2-Chloro-2'-deoxyadenosine (2CldA) is used for treatment of several lymphoid malignancies. Since this drug is incorporated into DNA, we have undertaken studies on base pairing of 2-chloroadenine (2C1A). 2CldA phosphoramidite was synthesized and used for preparation of 25-mer templates with 2C1A located at site 21 from the 3'-end. Kinetic parameters (Km and Vmax) for the incorporation of dNTPs by AMV reverse transcriptase opposite the 2C1A template, as well as for the extension of a 2C1A-T pair, were determined. The efficiency (Kin/Vmax) of incorporation of dGTP, dCTP and dATP opposite 2CIA is at least one order of magnitude lower than opposite an unmodified A.

43 The efficiency of incorporation of dTTP opposite 2C1A is 25 - 50-fold lower than opposite A and extension of an 2CIA-T pair is 3-fold lower than of an A-T pair. This indicates that the presence of 2CldA in DNA slows replication, but does not lead to base-substitution mutations. From the analysis of the parameters of dTTP incorporation we conclude that formation of a 2C1A-T pair is thermodynamically, but not kinetically controlled. The diffrence in binding energy (DDG) between 2C1AT and A-T pairs in the environment of the polymerase active site is 2 kcal/mol. 2'-Deoxyisoguanosine (diG, 2-oxo-2'deoxyadenosine) is formed in DNA in a reaction of 2'-deoxyadenosine with reactive oxygen species. In order to study its miscoding properties, 25-mer deoxyoligonucleotide templates with diG located site- specifically, were prepared.

2.2. Repair of cyclic adducts.

The combined action of glycosylases and abasic site-specific endonucleases on damaged bases in DNA results in single strand breaks. In plasmid DNA, as a consequence, the covalently closed circular (ccc) form is converted to the open circular (oc) form, and this can be quantitated by agarose gel electrophoresis. We studied DNA lesions sensitive to E. coli AlkA and cloned human ANPG-40 glycosylases which are known to excise alkylated bases and etheno adducts. To our surprise pBR322 and pAlklO plasmids not pretreated with mutagens were cleaved by both glycosylases in the presence of enzymes possessing endonucleolityc activity (Nth, Fpg, ExoIII), which indicates that plasmids contain unknown, endogenously formed adducts. Plasmid pretreated with chloroacetaldehyde, a mutagen forming etheno adducts, exhibited enhanced sensivity as expected. Adducts formed by acrolein and croton aldehyde are excised by AlkA, but not by ANPG-40, whereas maiondialdehyde adducts are not excise by either glycosylase. Bulky p-benzochinone adducts are not excised by AlkA, however, plasmid pretreated with this mutagen is incised by endonucleases, possibly without prior generation of an abasic site. These examples show that examination of conformational changes of plasmid DNA can be exploited to study the specificity of N- alkylpurine-DNA glycosylases. Publications: 3121, 1009/A, 1011/A PL0001647

3. PYRIMIDINE NUCLEOSIDE ANALOGUES, POTENTIAL CHEMOTHERAPEUTIC AGENTS, AND SUBSTRATES/INHIBITORS IN VARIOUS ENZYME SYSTEMS

T. Kulikowski, M. Bretner, K. Felczak, A. Drabikowska, and D. Shugar

Pyrimidine nucleoside analogues are an important class of compounds with antimetabolic (antitumor, antiparasitic and antiviral) properties. The synthesis of thiated nucleoside and nucleotide analogues, determination of structures, conformation and dissociation constants, their potential chemotherapeutic activities, and their

44 substrate/inhibitor properties in various enzyme systems, with emphasis on enzymes related to chemotherapeutic activities, were investigated. In the series of thionated inhibitors of thymidylate synthase (TS), potential antitumor agents, regioselective syntheses were elaborated for 2- and 4-thio, and 2,4- dithio derivatives of 2'-deoxyuridine (dUrd), 5-fluoro-2'-deoxyuridine (FdUrd), and several other 5-fluoro, 5-bromo- and 5-trifluoromethyl congeners, and the 2-thio derivatives of FdUrd and its oc-anomer, which proved to be selective agents with high cytotoxicities correlated with the inhibitory activities vs TS of their corresponding 5 '-monophosphates. Regioselective syntheses were also elaborated for 2'-deoxycytidine and 5-fluoro-2'-deoxycytidine derivatives. Solution conformations of these nucleosides were deduced from high- resolution (500 MHz) *H NMR spectra. Substrate/inhibitor properties of 2-thio-2'-deoxycytidine (S^dCyd) and 5-fluoro-2-thio-2'-deoxycytidine (S FdCyd) with respect to human leukemic spleen deoxycytidine kinase have been examined. Both are substrates, and also good inhibitors, of phosphorylation of 2'-deoxycytidine and 2'-deoxyadenosine. Particular attention was directed to the specificity of the NTP phosphate donor for several nucleoside kinases, and procedures have been developed for distinguishing between ATP and other NTP donors, a problem of importance in chemotherapy with nucleoside analogues. Biological properties of the newly synthesized thiated pyrimidine 2',3'- dideoxy-3'-fluoronucleosides, S^,3'-FddUrd and S^,3'-FddThd, were also investigated. Thiated 3'-fluoronucleosides were moderate substrates for thymidine phosphorylase and were quite inactive vs uridine phosphorylase. S ,3'-FddUrd proved to be a moderate, and S^,3'-FddThd a potent and selective, inhibitor of the replication of HIV-1 in CEM4 cells, comparable to the activity of the known antiretroviral agent AZT. 5-Fluorouridine and 5-fluorocytidine are potent antitumor agents, but too toxic to be used as drugs. To decrease this toxicity, a new class of branched-chain sugar nucleosides, pyrimidine and purine hamamelose nucleosides, was synthesized, with uracil and 5-fluorouracil derivatives being the most potent antileukemic agents, activities being comparable to that of 5-fiuorouridine. Publications: 3098, 3112,+3U3,3114, 3118,3137, 3138, 3139, 3142, 3150, 3175, 3182,3191, 3225, +3226,3236, 59/N

4. MECHANISMS OF PUVA (Psoralen + UVA) PHOTOCHEMOTHERAPY §=§

Z. Zarqbska, E. Waszkowska, D. Barszcz ^_ °j> ^= CD Our studies are aimed at understanding the action of PUVA phototherapy on := o the lym-phocytes membrane. The investigations are carried out on model lecithins ^= § having in the second position oleic/linoleic acid as the majority of lecithins have. a== =-* Previous investigations led to a discovery of a new class of photoadducts which ^= accompany the photolytic breakdown of lecithin. The new photoadducts were obtained =

45 by irradiating a mixture of psoralen or 8-methoxypsoralen with phosphatidylcholine; separated by HPLC or TLC and characterized by UV and NMR spectra and mass spectrometry. Kinetics of psoralen photolysis in the presence of lecithin was followed and the yield of the initial photoproducts was determined. The predominant reaction was a cycloaddition between one of the two photoreactive sites of psoralen (3,4-pyrone edge) and one of the olefinic bonds of the unsaturated fatty acid linked in the second position of the glycerol moiety of phosphatidylcholine. Two adducts were purified to homogeneity and characterized: an adduct of psoralen to oleic acid in lecithin (PCdlpal<>PSO) and two isomeric adducts to the free linoleic acid (d2<>PSO). The extreme sensitivity of isolated photoproducts was encountered in the course of identification analysis (UV, NR and mass spectrometry). This was in contrast to the other stable psoralen cyclobutane adducts, like psoralenothymine or psoralen<>fatty-acid-methyl-ester. We assume that the short life-time of our adducts is associated with the presence of a -COOH end group in the free fatty acid, and its ester group -COO- in the lecithins. These compounds undergo hydrolysis/oxidation reactions in aqueous micelles and in the slightly hydrated methanol. The photoadducts are also photosensitive, e.g. the cyclobutane ring splits under laser beam (337 ran, 50 mJ) applied in the MALD1 (Matrix Assisted Laser Desorption and Ionization) method for molecular mass estimation. With the more gentle method, LSIMS (Liquid Secondary Mass Spectrometry), molar ratio of compounds in photoadducts was found to be 1:1. Two M.Sc. Theses performed in our group contributed significantly to the progress of the investigations. The program is conducted in collaboration with the Department of Pharmaceutical Sciences of the Padua University, Centra di Studio sulla Chimica del Farmaco e dei Prodotti Biologicamente Attivi del CNR, Padova, Italy. Publications: 994/A, 997/A, 71/N, 72/N

^S 5. MECHANISMS OF UV-INDUCED MUTAGENESIS

——— _ /. Pietrzykowska, A. Bqbenek, A. Czajkowska, M. Felczak, J. Knvawicz

T- Studies on the isfA gene, presumably involved in the regulation of the switch-off of the i § SOS response, were continued. The influence of the isfA mutation on the specificity of 12 mutagenesis induced by UV-light and by alkylating agents was investigated in recA" i D- and recA730 E. coli strains. We have previously shown that the isfA mutation inhibits ! SOS-dependent phenomena, including SOS mutagenesis, most probably by inhibiting | RecA coprotease activity. UV-light induces mutations that are predominantly base substitutions, the largest group of which consists of the transitions AT-GC and GC-AT. Another mutagen which also induces GC-AT transitions, but by an SOS-independent pathway, is EMS. On the other hand, in the strain constitutively expressing the SOS functions, recA730, transversions are mainly observed. The effect of the isfA mutation on the mutation specificity induced by UV and EMS, or mutator activity in recA730, was analyzed in a system described by Cupples and Miller. It was found that the isfA

46 mutation inhibits the appearance of UV-induced GC-AT and AT-GC transitions in recA+ and GC-TA and AT-TA transversions in the recA730 mutator strain, but not GC- AT transitions induced by EMS. The results indicate that the isfA mutation affects the SOS-dependent mutagenic pathway, but has no effect on the specificity of induced mutations. Studies on the mechanism of UV-induced mutagenesis, which is independent of DNA replication in the lambda phage, were also continued. We have previously shown that UV-induced mutagenesis of the X susOn phage occurs under conditions nonpermissive for phage DNA replication in E. coli 594 su host, pointing to the existence of a new mutagenic pathway, which differs in several aspects from that occurring under conditions permissive for phage DNA replication. To learn more about the mechanism we compared the effect of a deficiency in mutHSL-dependent mismatch repair on UV- and MMS- induced mutagenesis of the X susOg phage under conditions nonpermissive or permissive for phage DNA replication. It was found that the mismatch repair + deficiency highly increases spontaneous and UV-induced reversion of X susO8 to sus in the C600 mutL host permissive, but not in the 594 sii mutL nonpermissive one, for phage DNA replication. The results further support the notion that mutations formed by a mechanism independent of DNA replication are not recognized by mismatch repair. A possible involvement of AP sites as premutagenic lesions in the DNA replication- independent pathway, as well as the effect of overproduction of UmuD', was studied. Overproduction of UmuD' increases MMS-induced mutagenesis more efficiently than that induced by UV, suggesting that UmuD' may be involved in mutagenic repair of AP-sites under conditions nonpermissive for DNA phage replication. Publications: 3103, 60N, 909/A, 910/A, 911/A, 912/A, 1006/A, 1007/A, 1052/A, 1053/A

47 DEPARTMENT OF PLANT BIOCHEMISTRY Head: professor Jerzy Buchowicz

Since 1995, when the latest issue of this series appeared, the Plant Biochemistry Department has continued its investigations on selected topics in plant molecular biology and biotechnology. Our attention was focused mainly on the occurrence of nuclear extrachromosomal DNA and its use in plant transformation, protein kinases and their role in signal transduction, and plant pathogenesis with a special reference to the increased resistance of transgenic potato to fungal and viral infections. The progress has not been, however, uniform in all directions. A considerable growth of knowledge resulted from studies on protein kinases in relation to environmental stresses. At least three protein phosphorylating enzymes that respond to salt stress were identified in tobacco. In collaboration with prof. J. Miernyk (USDA center in Peoria, IL, USA) a homologue of the bacterial heat shock protein hsp40 was isolated from the higher plant Arabidopsis thaliana. Protoplast fusion was used to obtain a potato callus tissue with an increased resistance to the pathogenic fungus Phytophthora infestans. Another approach, gene fusion, was successfully applied to detect a protein factor that influences the resistance of potato to the PVY virus. Very recently, attempts to prepare oral vaccines (against Helicobacter pylori) with the use of transgenic plants as bioreactors were undertaken.

1. CHROMOSOMAL AND EXTRACHROMOSOMAL DNA IN PLANT CELL DIFFERENTIATION

./. Buchowicz, B. Mazus, M. Dobrzanska, E. Kraszewska, B. Kroczynska, M. Bucholc, B. Sznrmak, C. Krysiak

A recently purchased device for biolistic transformation of intact cells (gene gun) was used to deliver foreign DNA into immature wheat embryos. The foreign DNA preparations were represented by constructs based on bacterial plasmids (pUC series) or nuclear extrachromosomal DNA of higher plants (reviewed in 3197). Sequences corresponding to the bacterial uidA and bar genes (reporter gene and marker gene, respectively) were inserted into each of the vectors. Conditions were found to achieve very efficient and reproducible transient expression of the reporter gene. The yield of stable transformation remained, however, relatively low (0.3%). Somatic embryogenesis, a stage necessary to regenerate transformed plants, was observed to limit the efficiency of stable transformation most severely. The morphogenetic capacity of explants was lowered by the biolistic bombardment to the extent that depended on the kind of the target tissue used. In particular, the frequency of embryogenic events was higher in bombarded callus tissues than in calli derived from bombarded embryos. Attempts to increase the yield of stable transformation are continued. Publications: +3158, 3164, 3197, 3198, 3209, 51/N, 1018/A

48 2. REVERSIBLE PHOSPHORYLATION OF PLANT PROTEINS

G. Muszynska, G. Dobrowolska, J. Szczegielniak, J. Trojanek, M. Mikolajczyk, I. Jurkowski, O. S. Awotunde

During the last two years the search for a calcium - phospholipid dependent protein kinase from maize seedlings and phosphorylation of tyrosine in plant proteins was continued. Additionaly, studies of subunit structure of plant protein phosphatase 2A and role of reversible phosphorylation in stress signalling were initiated. In order to identify protein kinases involved in a salinity stress response in plants, activities of several protein kinases were analyzed in cytosolic extracts of tobacco cells exposed to 250 raM NaCl. Under stress conditions activation of three protein kinases phosphorylating commonly used MAPKs substrates, myelin basic protein (MBP) and recombinant transcription factors c-Jun and ATF-2, was observed. One of the identified kinases can be assigned by immunological and biochemical criteria to the MAPK family, because it is recognized by an anti-MAPK antibody and its activity is regulated by dual phosphorylation on Thr and Tyr. Two other kinases do not resemble any of the so far known protein kinases. Moreover, in plants treated with high salt another unique protein kinase is activated. This kinase phosphorylates casein, but it differs significantly from typical casein kinases. All the identified kinases were activated transiently in the first minutes of stress application. Their activities are positiviely regulated by phosphorylation and they can be inactivated by maize protein phosphatase 2A. For the elucidation of the subunit structure of plant protein phosphatase 2A (PP2A) we elaborated the method of purification of the holoenzyme from maize seedlings to apparent homogeneity. SDS-PAGE of the holoenzyme has shown three protein bands of 65,55 and 36 kDa. The 65 kDa protein gives a positive reaction with antibodies against animal PP2A-65 kDa regulatory subunit. The 55 kDa protein was detected as another regulatory subunit by reactions with two types of antibodies raised against peptides from the N-terminal and the conserved region of the animal PP2A-55 kDa regulatory subunit. The 36 kDa protein was detected previously as the catalytic subunit. The results indicate the presence of a trimeric form of plant PP2A, consisting of one catalytic and two regulatory subunits. These data are the first biochemical evidence for the existence in plant PP2A of the 65 and 55 kDa regulatory subunits. Collaboration: dr. Stanislaw Zolnierowicz, Faculty of Biotechnology, Department of Biochemistry, Medical University of Gdansk. Publications: 3099,3160,3161,3234, 890A, 927A, 936A, 941 A, 942A, 943A, 944A, 1054A, 1055A, 1056A, 1057 A, 1070A, 1078A

49 3. TISSUE AND CELL CULTURES OF POTATO: PATHOGENS AND TRANSFORMATION

B. Wielgal, U. Maciejewska, A. Szczerbakowa, M. Borkowska, L. D. Was Hew ska, K. Kleczkowski

Our research concerns the resistance of potato to the pathogenic fungus Phytophthora infestans. The studies were conducted in two directions: 1) elucidation of biochemical mechanisms related to the resistance, 2) increasing the resistance of cultivated potato using biotechnology. All experiments were performed either with in vitro grown plants or cell suspension cultures. As elicitor, culture filtrate obtained from P. infestans was used. We focused our research on the oxidative burst, an early event in plant /pathogen interaction. The generation of active oxygen species (AOS), measured by luminol- dependent chemiluminescence (LDC), was examined in the elicitor treated cells of Solamtm tuberoswn cv. Tarpan and the dihaploid clone H8105, both susceptible to the pathogen, as well as in cells of the field resistant cv. Bzura and a completely resistant wild species S. nigrum. The Tarpan cells showed the highest and prolonged production of AOS in comparison with the others. In contrast, S. nigmm cells generated AOS in two phases, with peaks at 40 and 100 min after the elicitation. Exogenous catalase and ascorbate inhibited elicitor-induced LDC in all cells studied, whereas superoxide dismutase (SOD) increased LDC in cells of Bzura and S. nigrum but decreased in the Tarpan ones. Further studies on this phenomenon are in progress. The cvs Bzura and Tarpan were transformed with a soybean gene for 1,3-b glucanase. The PCR identification, hybridization, as well as an increased activity of 1,3-b glucanase confirmed the transfer of the soybean gene into potato genome in the case of cv. Bzura only. The transgenic plants, cultivated in vitro and in soil, showed limited increase in the resistance to P. infestans. Studies on fusion between protoplasts isolated from leaves of the wild species, resistant to P. infestans (S. nigrum, S. bulbocastanum) and those obtained from cell suspensions of S. tuberoswn dihaploid clone H8105 and cv. Bzura were undertaken.Various hybridization techniques "in tubes" and "in drops" were tested and a modification of the drop method, which yielded about 7% of heterofusion products, was chosen. Methods for selection of the putative hybrids and conditions for growth of parental and fused cells, as well as further plant regeneration are currently being examined. Collaboration: Department of Genetics, institute of Potato Research, Mtochow; Flow Cytometry Lab., Drug Institute, Warsaw. Publications: 3220, 973A, 907A, 1001A

50 4. MECHANISMS LEADING TO ACTIVATION OF PLANT DEFENSE GENES

J. Hennig, M. Koter, M. Krzymowska, A. Talarczyk

Previous studies argue that salycylic acid (SA) plays an important role in the plant signal transduction pathway(s) leading to disease resistance. It has been proposed that one of its modes of action is the inhibition of catalase and elevation of H2O2 level in the tissue. To verify the role of SA and H2O2 during pathogenesis, transgenic tobacco plants expressing Sacharomyces cerevisiae CTA1 gene coding for peroxisomal catalase were constructed. These plants possess 2-4-fold higher total catalase activity under normal growth conditions. No symptoms of chlorosis and/or necrosis were observed. After infection with tobacco mosaic virus (TMV), similar high levels of SA were detected in both CTA /-transgenic and control (Xanthi-nc) plants. However, levels of pathogenesis-related proteins (PR) and their respective mRNAs were significantly reduced in the infected leaves of the transgenic plants. No change in PR expression was detected in uninfected leaves of both CTA I and control plants challenged with TMV. These results suggest that elevation in catalase activity and the resulting reduction of HiO2 level results in more severe local disease symptoms, apparently due to an alteration of the hypersensitive response mechanism and does not influence systemic acquired resistance after viral infection. Regulation of the activity promoters of genes belonging to the PR family (PR- la, PR-2d), fused with the nidA gene (GUS) as a reporter, was tested individually in tobacco protoplasts. In contrast to the control (CaMV 35S promoter), marked differences in transient activity of both PR promoters were observed depending on the methylation state of the templates. High levels of GUS expression driven by PR-la or PR-2d were observed only when the template contained /Anethyl-deoxyadenosine in GATC sequences. Gel mobility shift assay was used to check for differences in binding of PR-la promoter fragments, containing GATC or GmATC sequences, with the tobacco nuclear extract. Presence of quantiative differences in the binding of the nuclear proteins to in vitro Dam methylated in comparison with non-methylated probes was detected. Our data suggest that 7V"methyl-deoxyadenosine, present in GATC sequences, may modulate the regulation of PR-s genes expression. Publications: 3115, +3154, 3155,3213, 36/N, 913/A, 974/A, 1002/A, 1003/A, 1077/A

5. ORAL VACCINES FROM PLANTS: CONSTRUCTION OF TRANSGENIC PLANTS PRODUCING RECOMBINANT UREASE OF HELICOBACTER PYLORI

A. Sirko, R. Brodzik, D. Gaganidze

Our aim is to use plants as bioreactors for the production of antigens of pathogenic bacteria or viruses. Transgenic plant tissues could be used for oral vaccination against respective pathogens, provided they are able to induce the mucosal

51 immunogenic response in animals. The most important advantage of this approach is a possibility of making very safe vaccines at a low cost. As a model antigen for our studies we have chosen urease of Helicobacter pylori, a Gram negative bacterium that colonizes human stomach. H. pylori is implicated in nearly all duodenal ulcers and most gastric ulcers and is associated with an increased risk of gastric adenocarcinoma. In developing countries 80% of the population is infected by age 20. Urease (consisting of two subunits) is the most prominent component of H. pylori and serves as a powerful immunogen. There is evidence that oral immunization with H. pylori urease (or its larger subunit) in the presence of adjuvants is a feasible strategy for development of such a vaccine. As a model plant we used low alcaloid line, LA Burley 21, of tobacco (Nicotiana tabacum) which is well tolerated by laboratory animals during immunological tests. We have constructed transgenic plants via Agrobacterhim- mediated transformation using binary plasmids containing expression cassettes with strong promoters governing transcription of the genes encoding subunits A and B of H.pylori urease (iireA and ureB, respectively). Eight out of over eighty screened tobacco plants transformed with the ureB cassette were producing UreB protein at high level. No plant producing UreA protein has been identified so far. The experiments leading to construction of transgenic plants producing both urease subunits are currently being conducted. Moreover, edible crops, like carrot (Daucus carrota) or pea {Pisnm sativum) which are more suitable for oral application will be transformed with the expression cassettes giving the best results in tobacco. Immunological tests on mice are planned to be performed in the closest future.

52 DEPARTMENT OF PROTEIN BIOSYNTESIS Head: professor Wiodzimierz Zagorski-Ostoja

The Department follows five main lines of research: (a) analysis of plant viroids and viruses structures and transformation of plants towards pathogen resistance; (b) genomic analysis of yeast; (c) regulation of gene expression in insect ontogenesis; (d) nucleolytic enzymes of higher plants and human carcinoma cells; (e) chromatin structures in transcriptional regulation. The first line of research concerns the pathogenicity of viroids and viruses in plants, viz. Potato Spindle Tuber Viroid (PSTVd), Potato Virus Y (PVY) and Potato Leafroll Virus (PLRV). The RNA pathogens studied cause considerable reduction in quality and yield of such important crops as potato, tomato, cucumber. Knowledge of the nucleotide sequences of viroid and virus genomes is crucial for the understanding of their pathogenicity, and allows us to construct plants (tobacco and potato) resistant to these pathogens. The second line of research focuses on functional yeast genome analysis. The significance of yeast as an experimental organism lies in the power of its genetics and its utility in biotechnology. The S.cerevisiae genome is relatively small, consisting of 16 chromosomes which have been completely sequenced. Our group participated in the European BRIDGE project sequencing fragments of chromosomes II, III, X and XL Functional analysis of the genes discovered by us is undertaken. 26 ORFs were inactivated and phenotypes of the deletant strains are under investigation. Progress in this field allows one to understand the functions of these new genes, their integration in the cell metabolism and evolutionary relations with genes of other organisms. Group working on the subject participates in the European Functional Analysis Program (EUROFAN). Regulation of gene expression in insects is the third problem in our studies. Investigations are focused on the role of receptor proteins in the transmission of hormonal signals. Special attention is given to the interaction between ecdysteroid receptor and its response elements. In studies on baculovims our objective is to characterize a new baculovirus SsMNPV isolate and to construct a recombinant baculovirus AcMNPV carrying synthetic poneratoxin gene which could be useful as a natural insecticide. Research on nucleolytic enzymes aims at their purification and characterization as probes for structural studies of RNA and DNA molecules. Two single-strand-specific nucleases, the Rn nuclease from rye germ nucleosol and the Chs nuclease from wheat chloroplasts, purified in our laboratory, have been employed to study secondary and tertiary structure of different tRNAs, 5S rRNAs, mRNAs and their complexes with proteins. A new, sequence-specific ribonuclease was purified from T84 colon carcinoma cell line. The potential utility of this RNase in the diagnosis of human colon carcinoma is under consideration. The fifth line of our studies concentrates on the regulation of chromatin structural transitions, probably one of the key mechanisms involved in the selective

53 transcription of genes during development. This work deals with proteins involved in ,,opening-up" of nucleosomes for transcription, and the mechanism of histone Hi- mediated transcriptional repression. Several nucleosome-disrupting proteins have been identified by sequence homology searches in the Arabidopsis data base, and their genes were cloned and are currently being analyzed. The mechanism of histone HI-mediated transcriptional repression is being studied in reconstituted chromatins containing natural DNA tracts showing different affinities for HI.

1. STRUCTURE OF POTATO SPINDLE TUBER VIROID (PSTVd) AND POTATO VIRUS Y (PVY) IN RELATION TO THEIR PATHOGENICITY IN PLANTS

W. Zagorski-Ostoja, P. Szafrafiski, A. Chachulska, A. Gora-Sochacka, A. Lipska- Dwuznik, J. Pawtowicz, M. Milner, W. Podstohki, E. Nowak, I. Rosa

Polymorhism analysis of phenotypically distinct local field isolates of potato virus Y (PVY) is analyzed. Full-length cDNA genome copies of selected isolates were synthesized and amplified. RFLP analysis of full genomes confirmed the polymorphism deduced from partial sequencing and enabled the localization of a polymorphic region, probably involved in the induction of necrosis in tobacco. Studies on the viral transcripts infectivity were carried on using different methods of plant infection. The sequence of the PVYN-Wi polymerase gene was established and conserved motifs were identified. Binary vectors carrying the PVY polymerase and NTR cDNAs were constructed in order to engineer plants resistant to PVY. Tobacco cv. Xanthi and SRI and potato cv. Irga were transformed by agroinfection and transgenic plants were regenerated. Several tobacco and potato lines, carrying a truncated PVY polymerase gene, highly or partially resistant to PVY were selected and subjected to further molecular analysis. Modeling of 3-D structures of viral proteins is actively followed. Work on the genetic stability of sequence variants of PSTVd was continued. A computer analysis of the secondary structure of the PSTVd RNA was carried out, leading to results on the thermodynamical stability of the pathogenicity (P) region. In order to construct PSTVd resistant plants, selected fragments of the PSTVd genomic cDNA were cloned in the binary vector pKYLX 71-35S", in (+) and (-) orientations. Potato plants (cv. Irga and Bintje) were transformed by agroinfection. Transgenic plants (analyzed by PCR) are being tested in the Plant Breeding and Acclimatization Institute in MIochow. Their analysis is continued using DNA and RNA hybridization methods. Publications: 3181, 3196, 3208, 3224, 3228, 3231, 3250,3251, 3252, 76/N, 999/A, 1028/A, 1032/A, 1058/A, 1059/A

54 2. SEQUENCING AND ANALYSIS OF THE YEAST GENOME

J. Rytka, W. Zagorski-Ostoja, M. Zagulski, R. Gromadka, R. Kucharczyk, A. Migdalski, B. Babinska, M. Ostrowska

Sequencing of a 4kb fragment from the right arm of chromosome X of the yeast S. cerevisiae strain S288C allowed us to identify several ORFs. One of them, YJR025c, shows interesting characteristics. The predicted protein sequence of yjrO25p shows significant similarity to the human 3-hydroxyanthranilic acid dioxygenase, an enzyme involved in tryptophan degradation and formation of nicotinic acid from tryptophan. Detailed genetic and biochemical analysis of the wild type and yjrO25c A:: HIS3 deleted strain proved that the YJR025c gene encodes 3-hydroxyanthranilic acid dioxygenase, an enzyme of unknown pathway of nicotinic acid synthesis in yeast. We demonstrated that the ORF YCL059c, discovered during the systematic sequencing of chromosome III, codes for a protein essential for cell growth. It contains a 131-aa fragment with a characteristic basic motif KRR-R highly conserved in the evolutionary distant species such as nematodes, plants and humans. We called this essential gene KRR1. The function of this gene is studied by means of classical and reverse genetics. Within the frame of EUROFAN network for functional analysis of the yeast genome we undertook systematic studies on new ORFs from chromosome II seqenced in our laboratory. The ORFs YBR128c, YBR I3Iw, YBR 133c, YBR 137w, YBR 138c and YBR142w were disrupted in two reference diploid strains by PCR targeting technique using kcmMX4 cassetes. The correct gene replacements were verified by PCR in the heterozygous diploid ORF A::KanMX4 and the four spore clones. The wild type genes were cloned by the gap-repair procedure. We have started the phenotypic analysis of heterozygous and homozygous diploids bearing null alleles of the ORFs studied and of the haploid spore clones. Collaboration: prof. P.P.SIonimski, Centre Genetique Moleculaire, CNRS, Gif-sur- Yvette, FRANCE Publications: 3125, 3128, 3135,1025/A, 1026/A, 1027/A

3. ROLE OF HORMONES IN GENE EXPRESSION OF Galleria mellonella

Z. Lassota, K. Grzelak, B. Kludkiewicz, K. Gocman

Studies on the influence of cold stress on the synthesis of Galleria mellonella proteins of mol. wt. from ca 70kD to 82kDa were continued (in cooperation with the Department of Invertebrate Physiology, Warsaw University). It was found that in Galleria larvae exposed to low temperature the synthesis of 74 kDa and 76 kDa proteins considerably precedes the synthesis of protein(s) of about 82 kDa. One of the cold shock proteins was identified. The protein of mol. wt. 82 kDa belongs to the LHP proteins class. Our results suggest that ecdysteroids act as negative regulators of the expression of cold shock protein genes.

55 Within a project carried out with the Biochemistry Department of the Technical University of Wroclaw our group started to develop a yeast (Pichia pastoris) expression system which would enable the production of recombinant proteins constituting elements of the functional ecdysteroid receptor. Collaboration: prof. Bronislaw Cymborowski, Jakub Godlewski, M. Sc, - Warsaw University; prof. M. Kochman, dr. A. Ozyhar - Technical University of Wroclaw. Publications: 3178, 920/A, 967/A, 1051/A, 54/N

4. BACULOVIRUSES AND THEIR RECOMBINANTS

I Z Lassota, \ J. Michalik, E. Szoiajska

Baculoviruses infecting insects can be used as ecologically safe and specific biopesticides. However, the characteristics of their genome as well as species - specific virulence in mixed populations have to be established before releasing these baculoviruses in to the environment. We are trying to find an appropriate insect cell line(s) supporting SsMNPV virus isolate replication. Baculovirus from Stilpnotia salicis does not infect any of the commonly used insect cell lines. For this reason a new insect cell line from the natural host of SsMNPV baculovirus will be established. Currently work on physical map of the SsMNPV isolate genome is under way. Publications: 3186, 3229, 1033/A, 1034/A

5. NUCLEOLYTIC ENZYMES FROM HIGHER PLANTS AND HUMAN CARCINOMA CELLS

J. W. Szarkowski, E. Killigowska, A. Przykorskct, M. A. Shvecka, hi. Tomaszewski, G. Przewlocki, D. Klarkowska

Identity elements in E.coli tRNACys were investigated using enzymatic methods. We have shown that cysteinyl tRNA synthetase protects the anticodon loop region of its cognate tRNA from digestion by single strand specific nuclease Chs at positions 34, 35, 36, 37 and the acceptor stem at position 73. This confirms that the anticodon loop and position 73 in the acceptor stem are the major identity elements in E.coli tRNACys. Another region in tRNACys protected by the synthetase from double- strand specific nuclease VI digestion is the acceptor stem region at positions 6, 7 and 65, 66. In contrast, position 64 and positions 51, 52, 53 in the T^P stem are better accesible for the nuclease in a complex with the synthetase than in free tRNACys, suggesting a structure change of tRNACys upon synthetase binding. Studies on ribosomal nuclease II were continued. The nuclease II does not hydrolyze synthetic analogs of cap structures of mRNA, methylated and unmethylated 7 7 (m Gp3A, GP3A and m Gp3C). The enzyme recognizes C-A and U-A bonds, both in

56 double-stranded and loop regions. This enzyme may serve as a probe in investigations on structures of nucleic acids. New RNase T84 isolated from human colon carcinoma T84 cell line, was shown to be free of other RNase activities. This enzyme has a molecular weight of 19 kDa and cleaves RNA in a sequence-specific manner, and the site of cleavage is located at the 3' side of A or G and the 5' side of U. RNase T84 degrades RNA chains to 3-OH and 5 -phosphate-terminated fragments. Publications: 3202, 996/A, 1029/A

6. IDENTIFICATION OF ARABIDOPSIS GENES INVOLVED IN NUCLEOSOME DISRUPTION

A. Jerzmanowski, J. Brzeski, T. Calikowski, P. Koibiai, M. Przewloka, M. Prymakowska-Bosak, R. Tomaszewski (mixedgroup: IBB PAN, Department of Biology, Warsaw University)

In the early 1980s Herskowitz and colleagues discovered that mutations in a set of ,,Switch" and ,,SNF" genes in yeast reduce expression of a number of inducible genes by depriving the cell of a system for active removal of nucleosomes. More recent experiments have shown that the products of Drosophila genes involved in early development, mammalian steroid receptors and the yeast transcription factor GAL4, all stimulate transcription through mechanisms dependent on SWI/SNF like activities. The function of the SWI/SNF complex seems to be conserved in eukaryotes. Using the Arabidopsis thaliana EST (expressed sequence tags) clones, we have identified an Arabidopsis homolog of the yeast SNF 5 gene and obtained a complete sequence of its cDNA. With 240 amino acids the Arabidopsis homolg of yeast SNF5 is the smallest SNF5-type protein identified so far. Unlike yeast SNF5 and its human homolg INI1, the Arabidopsis protein is unable to activate transcription in yeast when tethered to DNA. We showed that mRNA of the Arabidopsis SNF-5 protein is ubiquitously expressed in the plant. Transgenic Arabidopsis plants with markedly decreased level of SNF5-type, mRNA are viable but exhibit a distinctive phenotype characterized by bushy growth and flowers that are mostly unable to produce seeds. Publications: 1046/A

7. HISTONE HI-MEDIATED TRANSCRIPTIONAL REPRESSION

A. Jerzmanowski, J. Brzeski, T. Calikowski, P. Kozbial, M. Przewloka, M. Prymakowska-Bosak, R. Tomaszewski (mixed group: IBB PAN, Department of Biology, Warsaw University)

Transgenic tobacco plants with modified expression of homologous and heterologous genes of histone HI have been constructed. Transgenic plants have been selected with markedly aberrant stoichiometry of histone H1:DNA as well as with

57 changed profile of main chromosomal variants of histone HI. The effects of these changes on phenotypic and ultrastructural appearance, transcriptional profile and structure of chromatin is being analyzed. In order to study the function of histone HI phosphorylation, transgenic tobacco BY2 cells were constructed that bear the Arabidopsis thaliana HI gene with main phosphorylation sites knocked-out by Ser/Thr^Ala replacement. Histone HI plays an active role in establishing the transcriptionally repressed chromatin state of the oocyte-type 5S RNA genes in the early stages of Xenopus development. By using a fully defined in vitro system of chromatin assembly, we found that the DNA fragment comprising the oocyte-type 5S RNA genes with its native flanks (but not the somatic-type 5S RNA gene with its GC-rich flanks) is capable of directing the histone HI-mediated chromatin reorganization of densely packed DNAxore histone complexes on long stretches of DNA. The reorganization results in creation of regular long gaps between packed core particles and is achieved through complete removal of part of the core histone complexes from DNA template. The binding of HI results in protection of DNA sites within the AT-rich oocyte-type repeat and in spaced nucleosomes that are partly stabilized at specific positions over template DNA. These results support the view that the AT-rich flanks of the oocyte-type 5S RNA gene can be the major cause of histone HI-mediated chromatin reorganization that results in transcriptional repression observed in vivo. Work is in progress on the elucidation of the molecular mechanisms of HI action in the above system. Publications: 3146, 3179, 3199, 1044/A, 1045/A, 1047/A,1048/A, 1049/A, 1050/A

58 LABORATORY OF MUTAGENESIS AND DNA REPAIR Head: professor Zygmunt Ciesla

Two main lines of research concern the mechanisms controlling fidelity of DNA replication in Escherichia coli and regulation of cellular responses of Saccharomyces cerevisiae to DNA damaging agents. The mechanisms that control the fidelity of DNA replication are a matter of considerable interest. These mechanisms are being investigated by a number of approaches, including mutagenesis, detailed kinetic and structural studies. We investigate mutagenic potentials of leading and lagging strands during DNA synthesis in the model organism Escherichia coli. In order to address this question, we constructed a new system that allows the examination of mutagenesis of the same target gene in two orientations with respect to the origin of DNA replication, such that a given DNA sequence would be replicated as a leading strand in one orientation and as a lagging strand in the other. We are also interested in elucidating the physiological role of interactions between the a, e and 9 subunits in the DNA polymerase III core. In an attempt to achieve this goal, we use the Saccharomyces cerevisiae two-hybrid system. Using this system, we investigate in vivo interactions between wild-type and mutant DnaQ proteins with the wild-type DnaE and HolE proteins. In response to DNA damage eukaryotic cells arrest the cell cycle and induce transcription of genes that facilitate DNA repair processes. Pathways that control these responses are being explored in several organisms. One approach to study cellular responses of S. cerevisiae to DNA damaging agents was based on characterization of novel DIN (damage inducible) genes. Main effort was concentrated on the characterization of the DIN7 gene. The product of DIN7 belongs to a large family of proteins including Rad2, Rad27 and Exol of S. cerevisiae and their human homologues. All these proteins are endowed with DNA nuclease activity. Persistent efforts to identify a physiological function of Din7 resulted in finding that the protein is involved in maintaining stability of mitochondrial DNA. In another project, the reading frame YBR173c on the chromosome II of S. cerevisiae was identified as a novel DNA damage inducible gene - DIN8 Another project which concerns the regulation of S. cerevisie response to DNA damage involved studies on the expression of the MAGI gene coding for 3- methyladenine glycosylase. Activity of this important DNA-repair enzyme of broad substrate specificity is induced in cells treated with various DNA-damaging agents. The MAG! gene has been classified as a DIN gene. Our investigations have shown that the products of the known regulatory genes engaged in the signal transduction pathway from DNA damage to the promoters of DIN genes, MECI, RAD53 and DUNI, are not required for transcriptional activation of MAGI.

59 1. REGULATION OF THE FIDELITY OF DNA REPLICATION IN ESCHER1CHIA COU

I. Fijalkowska, P. Jonczyk, M. Maliszewska-Tkaczyk, D. Gawel, M. Bialoskorska

During chromosomal DNA replication, the two DNA strands are replicated asymmetrically, one strand (the leading strand) continuously, the other (the lagging strand) discontinuously in the form of Okazaki fragments. The different modes of replication present the possibility that the two strands are copied with different fidelities. We investigated this during in vivo replication of the Escherichia coli chromosome by measuring lacZ reversion frequencies in strains containing the lac operon in two different orientations with respect to the chromosomal replication origin. To measure the specificity and frequency of mutagenesis a set of lacZ alleles constructed by Cupples and Miller (1989) were used. Our results obtained with mutator strains impaired in mismatch repair or in proofreading activity indicate that there is a difference in the fidelity of replication between the two replicating DNA strands. The differences in the level of mutagenesis between the two strands were observed both, for transitions and transversions. In contrast to previous conclusions from plasmid-based systems, on the E. coli chromosome lagging-strand replication may be more accurate than leading-strand replication. A mechanism coordinating DNA polymerization and DNA excision, relying on structural and functional communication between different subunits of Pol III holoenzyme, may play an important role in maintaining optimal fidelity of DNA replication. We used the Saccharomyces cerevisiae two-hybrid system to analyze specific in vivo protein-protein interactions between E. coli Pol III core subunits. We were particularly interested in elucidating the physiological role of interactions between DnaE (a subunit - possessing DNA polymerase activity) and DnaQ (s subunit - possessing 3'-5' exonuclease activity). We tested the interaction of the wild-type DnaE and HolE proteins with three well known mutant forms of DnaQ (MutD5, DnaQ926 and DnaQ49), each of which leads to a strong mutator phenotype. Our results show that the mulD5 and dnaQ926 mutations do not affect the s-a and e--6 interactions. However, the dnaQ49 mutation greatly reduces the strength of interaction of the s subunit with both the a and the 6 subunit. Thus, the mutator phenotype of dnaQ49 may be the result of an altered protein conformation of the e protein which, as a consequence, leads to altered interactions within the pol III core. Publications: 3123,+3141, 3221,+3241, 77/N, 1005/A, 1013/A, 1017/A, 1021/A, 1085/A, 1086/A

60 PL0001650

2. CELLULAR RESPONSES OF SACCHAROMYCES CEREVISIAE TO DNA DAMAGE

Z Ciesla, E. Sledziewska-Gojska, A. Nowicka, P. Mieczkowski, M. U. Fikus, P. Koprowski

Several experimental strategies have been used to study responses of S. cerevisiae cells to DNA damage. One approach was based on the isolation of novel genes, the expression of which is induced by lesions in DNA. One of these genes, DIN7, was cloned and partially characterized previously. The product of D1N7 belongs to a large family of proteins involved in DNA repair and mutagenesis. This family includes Rad2, Rad27 and Exol proteins of S. cerevisiae and their respective human homologues, all of which are endowed with DNA nuclease activity.To study cellular function of Din7 we constructed the pPK3 plasmid carrying DIN7 fused to the GAL1 promoter. Effects of Din7 overproduction on the phenotypes of wild-type cells and of rad27 and exol mutants were examined. Overproduction of Din7 does not seem to affect the proficiency of wild-type S. cerevisiae cells in recombination and mutagenesis. Also, overexpression of DIN7 does not suppress the deficiency of the EXOI gene product, the closest homologue of Din7, both in recombination and in controlling the fidelity of DNA replication . Unexpectedly, we found that elevated levels of Din7 result in a very high frequency of mitochondrial rhd mutants. A high frequency of production of rho mutants was also observed in strains defective in the functioning of the Dunl protein kinase involved in signal transmission in cells exposed to DNA damaging agents. Interestingly, deficiency of Dunl results also in a significant derepression of the DIN7 gene. Experiments are under way to distinguish whether a high cellular level of Din7 specifically decreases stability of mitochondrial DNA or affects stability of chromosomal DNA as well. Analysis of previously constructed S. cerevisiae strains carrying random genomic fusions with reporter lacZ gene, allowed us to identify the reading frame YBR173c, on chromosome II as a novel damage inducible gene - DIN8 . We have shown that DIN8-lacZ fusion is induced in yeast cells treated with MMS or exposed to UV light. Northern RNA analysis indicates that DIN8 is induced in response to DNA damage at the transcriptional level. DINS was cloned and the phenotype of cells with disruption of the gene is under study. POL2-MECI-RAD53-DUNI-signa\ transducing pathway has recently been postulated to be involved in the regulation of response of S. cerevisiae cells to DNA- damaging agents. We analyzed the expression of a known damage inducible DNA- repair gene, MAGI, encoding 3-methyladenine glycosylase, in S. cerevisiae strains carrying MAGlr.lacZ fusion and deficient in either POL2, MEC1, RAD53 or DUN1 function. P-galactosidase activity was assayed in cycling cells exposed to MMS or UV light. It was found that, in contrast to model DNA damage inducible RNR genes, neither mutation in the sensory C-terminal part of polymerase e (pol2-l 1) nor the in the Mecl, Sadl/Rad53 or Dunl cellular kinases blocks the induction of MAGI in response to MMS or UV light in cycling yeasts.

61 Publications: 3123, 3188, 3189, 3143, 966/A, 968/A, 1010/A, 1012/A, 1019/A, 1083/A, 1093/A, 80/N

LABORATORY OF PURINE METABOLISM Head: professor Maria M. Jezewska

PROPERTIES OF SOME ENZYMES OF PURINE METABOLISM M. M. Jezewska, H. Trembacz and G. Jewdoszuk

Our Laboratory is engaged in comparative studies of purine metabolism in various organisms. Two adenosine phosphorylases previously found in the trematode Fasciola hepatica, the enzymes not occurring in the vertebrate hosts of parasites, were investigated. Recent studies concerned the kinetics, activity towards various Ado analogues and the inhibitory effects thereof. The Ado phosphorylase which cleaved Ado, 2 -dAdo and 5 -methylthioadenosine (MTA) appeared to be similar to the Ado/MTA phosphorylase of parasitic protozoan Trypanosoma cruzei. The other Ado phosphorylase of F. hepatica resembles that found in the snail H. pomatia and described by us previously. The kinetics of F. hepatica enzyme is simpler than that of purine nucleoside phosphorylases known to date. This enzyme is strictly specific towards Ado and 2 - dAdo. However, several Ado analogues modified in the ribose moiety exert a weak inhibitory effects on it, and analogues modified in the purine ring are stronger inhibitors. The metabolic importance of this Ado phosphorylase for F. hepatica is not yet established. Possibly, this enzyme could be a target for a fascioliasis chemotherapy non-interfering with the host metabolism of Ado and MTA. The enzymatic preparations obtained by us from F. hepatica and H. pomatia not only cleaved MTA phosphorolytically but also deaminated it. The MTA deamination rate is comparable to those of Ado and 2 -, 3 - and 5 - deoxy analogues of Ado. For all these substrates the Michaelis constants of H. pomatia enzyme are within the range of 16 - 60 mM. The inablility to deaminate adenine, AMP and S- adenosylhomocysteine suggests that the enzyme is an Ado deaminase of broad substrate specificity. A deaminating activity towards MTA has not been reported to occur in vertebrates. Its presence in F. hepatica draws attention to a possibility of the deamination of Ado analoques, seen by many as potential drugs against trematode parasites. Publications: 3205, 924/A, 983/A, 995/A

62 LABORATORY OF BIOLOGICAL NMR Head: professor Andrzej J. Bierzynski

The objective of the Laboratory is to provide access to a high-resolution NMR facility, as well as scientific and technical assistance for scientists from all Polish laboratories involved in structural studies of biologically important molecules. The laboratory is equipped with a 500-MHz Varian UnityPlus instrument with three channels, ultrashim system, digital processing, and two - 5 mm and 8 mm - triple 'H/ljC/I5N probes with gradients. Full hardware and software computer equipment for analysis of 1, 2, and 3D spectra, structure simulations, and molecular dynamics calculations is also available. The following projects are being pursued:

Investigation object Main investigator Affiliation

Calcium binding peptides A. Bierzynski Inst. of Biochem. and Biophys. PAS CMTII and its M8-L A. Bierzynski Inst. of Biochem. and Biophys. PAS mutant S100A1 protein A. Ejchart Inst. of Biochem. and Biophys. PAS Cerebrospinal fluid B. Toczylowska Inst. of Biochem. and Biophys. PAS Psoralen photoadducts Z. Zar?bska Inst. of Biochem. and Biophys. PAS BPTI mutants M. Dadlez Inst. of Biochem. and Biophys. PAS Cyclodextrin-camphor H. Dodziuk Inst. of Organic Chemistry PAS system Nucleotide derivatives R. Stolarski Warsaw University mRNA-protein/peptide R. Stolarski Warsaw University systems Nucleotide-protein M. Remin Warsaw University interactions Extracts of plant tissues K.. Kaminska-Trela Inst. of Organic Chemistry PAS Scyliorhinin I] K. Rolka Gdansk University AMCI protein and BPTI J. Otlewski Wroclaw University mutants Protamine W. Bal Wroclaw University Sugars-ions interactions S. Tyrlik Agricultural and Pedagogical University, Siedlce Extracts of cancer tissues M. Mossakowski Institute-Centre of Experimental Medicine PAS

Publications: 3212,3216,3222,3245,1093/A, 1094/A

63 BIOINFORMATICS UNIT Head: Associate professor Piotr Zielenkiewicz

P. Zielenkiewicz, A. Kierzek, D. Plochocka, S. Kaczanowski

The Bioinformatics Unit of the Institute runs the Polish National Node of the European Molecular Network (EMBnet), which is a European non-profit organization for data access and distribution of information and software in the fields of molecular biology, genetics and biotechnology. It is composed of a network of nationally mandated and special nodes, each being a recognized biocomputing centre. The Polish National Node located at IBB was authorized by the State Committee for Scientific Research (KBN) and established in autumn 1994. From fall 1997 the bioinformatics node at IBB serves also as a Regional Node of UNESCO's ICCBnet (International Center for Cooperation in Bioinformatics network). The latest releases of nucleotide (EMBL - updated weekly and GenBank) and protein (SWISSPROT, PIR and TREMBL) databases are available locally, together with a restriction enzyme database (REBASE) and a collection of protein sites (PROSITE). The Genetics Computer Group (GCG) package of over 100 computer programs for sequence manipulation, database searching, sequence analysis and structure prediction is the most widely used software. Software tools for molecular evolution studies are available (PHYLIP, CLUSTALV), as well as programs for analyzing genomic information (ACEDB). Searching remote databases is made possible through links to the relevant sites on the World Wide Web (e.g. Software Retrieval System SRS), and for clients qf the BLAST and "entrez" services at the National Centr for Biotechnology Information (USA). Molecular modeling studies are possible with the use of commercial packages of BIOSYM/MSI and TRIPOS Ass. The node currently serves a community of around 600 users from academic institutions in Poland. User support and training is provided, including bioinformatics courses, both national and international, organized at IBB. The responsibilities of the Unit involve also bioinformatics infrastructure planning and coordination and maintenance of IBB's World Wide Web page at http://www.ibb.waw.pl which can be readily consulted with regard to current activities of the Institute and the Unit. The research activities of the Unit concentrate on the development of new algorithms for sequence comparison and biopolymer structure analysis.

64 LABORATORY OF MOLECULAR BIOLOGY (affiliated with the University of Gdansk)

1. MOLECULAR MECHANISMS OF ASSEMBLY, INHERITANCE AND DISASSEMBLY OF BACTERIOPHAGE A, REPLICATION COMPLEX

| K. Taylor , \ A. Wqgrzyn, G. Wqgrzyn

We have found that, in contrast to the free O initiator protein of X phage or plasmid rapidly degraded by the Escherichia coli ClpP/ClpX protease, the XO present in the replication complex (RC) is protected from proteolysis. RC is inherited by one of the two daughter X plasmid copies after each replication round and can function for many cell generations. However, in cells growing in a complete medium a temperature shift from 30 to 43°C resulted in the decay of this XO fraction, which indicated disassembly of RC. This process occurred due to heat shock induction of the groE operon, coding for molecular chaperones of the Hsp60 system. We demonstrated that a mere increase in cellular concentration of GroEL and GroES proteins is not sufficient to cause RC disassembly. Another requirement is a DNA gyrase-mediated negative re-supercoiling of X plasmid DNA which counteracts DNA relaxation and starts to dominate 10 min after the temperature upshift. We presume that RC dissociates from X DNA during the negative re-supercoiling, becoming prone to the subsequent action of GroEL/S and ClpP/ClpX proteins. Contrary to Xcro+, in Xcro' plasmid-harboring cells the RC reveals heat shock resistance. After temperature upshift of the Xcrots plasmid-harboring cells, a Cro repressor-independent control of X DNA replication and heat shock resistance of RC are established before the period of DNA gyrase-mediated negative supercoiling. We suggest that the tight binding of RC to X DNA is due to the interaction of RC with other DNA-bound proteins, and is related to the molecular basis of the Xcro' plasmid replication control. Publications: 3166, 3167, 3168, 3169, 3247, 3248, 3249, 325S, 3256, 915/A, 916/A, 918/A, 919/A, 926/A, 1079/A, 1080/A, 1095/A, 1096/A, 1097/A, 1098/A, 1099/A, 1100/A, 1101/A, 1102/A

: Department of Molecular Biology, University of Gdansk

65 2. MECHANISMS OF FtsH - DEPENDENT DEGRADATION OF THE a32 TRANSCRIPTIONAL REGULATOR OF ESCHERICHIA COLI AND THE ROLE OF CHAPERONE PROTEINS IN THIS PROCESS

A. Blaszczak, K. Liberek, M. Zylicz

The E. coli a32 transcriptional regulator has been shown to be degraded both in vivo and in vitro by the FtsH (HflB) protease, a member of the AAA protein family, [n our attempts to study this process in detail we found that two a32 mutants lacking the 15-20 C-terminal amino acids had substantially increased half-lifes in vivo and in vitro, as compared to the wild type a32. A truncated version of a32, CJ32CA, was purified to homogeneity and shown to be resistant to FtsH-dependent degradation in vitro, suggesting that FtsH initiates o32 degradation from its extreme carboxyl terminal region. Purified G32CA interacted with the DnaK and DnaJ chaperone proteins in a fashion similar to that of the wild type G32. However, in contrast to the wild type a32,

*: Department of Molecular and Cellular Biology, University of Gdansk

66 RESEARCH UNITS OF THE WARSAW UNIVERSITY LOCATED AT THE INSTITUTE

NEXT PAGE(S) left BLANK DEPARTMENT OF GENETICS (associated with the Institute) Head: professor Piotr Wegleñski

1. MOLECULAR ANALYSIS OF GENES CONTROLING ARGININE CATABOLISM IN ASPERGILLUS NIDULANS

P. Wçglenski, A. Borsuk, A. Dzikowska, J. Empel, A. Grzelak, J. Klimczuk, A. Tomczyk, J. Szyszka, M. Tolak

The main regulatory gene (arcA) controling expression of the arginine catabolic enzymes was cloned and sequenced. The gene codes for the protein containing the binuclear zinc finger, typical for many fungal transcriptional activators. The arcA gene was also sequenced in the arcA47 mutant. This mutation leads to the high constitutive synthesis of the arginine catabolic enzymes. Mutation was found to be the single nucleotide substitution resulting in the replacement of leucine by isoleucine within the first loop of the zinc finger. Analysis of the promoter of the agaA gene coding for arginase revealed several interesting structural properties, indicating a possibility of autocatalytic processing of mRNA and of direct involvement of arginine in this process. The checking of these hypotheses in under way. Publications: 3204

2. MOLECULAR MECHANISMS OF RNA PROCESSING IN YEAST MITOCHONDRIA

P. Stçpien, E. Bartnik, A. Dmochowska, P. Golik

Our research is aimed at elucidating the functions of several nuclear genes which have a major impact on mitochondria! RNA metabolism. One of these genes is SUV3, which was discovered as a nuclear suppressor of a deletion in the mitochondrial vari gene. This mutant displays a very complex phenotype: it has pleiotropic effects on mitochondrial mRNA processing, leads to accumulation of excised introns, changes 3' end processing of RNA and causes differences in levels of some mRNAs. Recently we have been able to show that the SUV3 protein is a splicing factor for the groupl intron rl (omega) from the mitochondrial 21SrRNA gene. We have constructed a yeast mt genome containing only one intron, rl, and introduced this genome into a strain bearing disrupted SUV3. The presence of the rl intron resulted in a block in respiration, change in the splicing pattern of the excised intron and a decrease in the amount of 2IS rRNA. We assume that so-called self-splicing introns require multicomponent protein-RNA complexes for the in vivo splicing reactions.

69 We have cloned and sequenced the previously unknown yeast nuclear gene DSS- 1, whose product is probably involved in regulating mRNA stability. We have also discovered that the SUV3 protein regulates mt mRNA stability in an intron-dependent manner. We have isolated four additional yeast genes whose products possibly inteeract with the suv3p and we investigate the role of these proteins in mitochondrial RNA metabolism. Collaboration: P. Slonimski, J. Lazowska - Centre Genetique Moleculaire CNRS, Gif-sur-Yvette, France. Publications: 3076, 3077, 3080

70 INSTITUTE OF EXPERIMENTAL PLANT B1O1 X)C.Y Director : professor Andrzej Podstolski

DEPARTMENT OF PLANT BIOENERGETICS Head: professor Anna Rychter

REGULATION OF PLANT ENERGY ECONOMY BY ENVIRONMENTAL CONDITIONS

A. Rychter, I. Ciereszko, A. Gniazdowska, I. Juszczuk, E. Malusa, M. Mikulska, A. Kondracka, M. Wanke

Phosphate deficiency in growth medium induces modifications in the ATP producing and utilizing processes. In leaves, low phosphate level affects the rate of photosynthesis only slightly but induces changes in photorespiratory metabolism and other metabolic routes regulating intracellular orthophosphate level. Both in leaves and in roots lower ATP and higher NAD(P)H levels are observed. The adverse energy balance may modify plasma membrane properties and thus nitrate uptake. Lower ATP level in leaves and roots is the result of increased participation of the alternative respiratory pathway. The alternative pathway may be induced by increased free radical formation. The localization, metabolism and physiological role of sugar accumulation in phosphate deficient roots was also studied. In the studies of regulatory mechanisms controlling the transcription of C4 PEPC genes our results support the hypothesis of a negative regulation of SVC4 expression that is modulated by phosphorylation using phytochrome as the photoreceptor. Publications: Ciereszko I., Gniazdowska A., Mikulska M. and Rychter A. 1996. Assimilate translocation in bean plants {Phaseolus vulgaris L.) during phosphate deficiency. J. Plant. Physiol. 149, 343-348. Kondracka A. and Rychter A. 1997. The role of Pi recycling processes during photosynthesis in phosphate deficient bean plants. J. Exp. Botany, 48, 1461-1468 Juszczuk I. M. and Rychter A. 1997. The changes in pyridine nucleotide levels in leaves and roots of bean plants (Phaseolus vulgaris L.) during phosphate deficiency. J. Plant Physiol. Hi, 399-404 Malusa E. and Rychter A. 1997. Effect of phosphate deficiency on isoenzyme pattern of hydrogen scavenging enzymes in bean roots. Proceedings of XIV National Congress of the Italian Society of Agrarian Chemistry-SICA, Rimini 1996 (427-433). Rychter A. M. 1996. The plant respiratory chain. Post. Biochem. 42, 268-276 (in Polish)

71 DEPARTMENT OF PLANT RESISTANCE Head: professor Alina Kacperska

PLANT RESISTANCE TO ABIOTIC STRESSES: PHYSIOLOGICAL AND BIOCHEMICAL MECHANISMS INVOLVED IN PLANT ACCLIMATION TO LOW TEMPERATURE AND DEHYDRATION ON MOLECULAR, CELLULAR, TISSUES/ORGAN AND WHOLE PLANT LEVELS

A. Kacperska, S. Egierszdorff, M. Kubacka-Zebalska, D. Solecka, J. Spychalska

Cold- and frost-induced modifications in the structure and chemical composition of cell walls in leaves of winter oilseed rape (Brassica napus. L, var. oleifera L) were characterized. Acclimation of plants in cold (>0°C) resulted in a marked increase of cell wall content and thickness due to the accumulation of pectic polysaccharides. Sugar composition of pectins was also modified. Transient freezing (-5°C for 18 h), giving rise to degradation of pectins, activated transcription of extensin genes (experiments performed by Piotr Kozbial of the Plant Mol. Biol. Lab). Low temperature during the postfreezing period promoted accumulation of extensin mRNAs. The role of the cold- and frost-induced cell wall modifications in plant adjustment to low temperature-induced water stress was poined out. Using nuclear run on technique and hybridization probes (selected EST clones from Arabidopsis thaliana), cold- and frost-dependent changes in transcription of several genes were described (Piotr KozbiaJ and Daniel Pukacki of the Plant Mol. Biol. Lab). Publications: Kacperska A. (1997) Ethylene synthesis and a role in plant responses to different stressors. In: Biology and Biotechnology of the Plant Hormone, Ethylene. (A. A. Kanellis, eds) pp: 207-216, Kluwer Academic Publishers, the Netherland. Solecka D (1997) Role of phenylpropanoid compounds in plant responses to different stress factors. Acta Physiol. Plant. !£, 257-268.

PLANT MOLECULAR BIOLOGY LABORATORY

Head: professor Andrzej Jerzmanowski

HISTONE HI IN TRANSCRIPTIONAL REGULATION

A. Jerzmanowski, M. Prymakowska-Bosak, M, Przewloka, P. Koibial Transgenic tobacco plants with modified expression of homologous and heterologous genes of histone HI have been constructed. Transgenic plants have been selected with markedly aberrant stoichiometry of histone H1:DNA as well as with changed profile of main chromosomal variants of histone HI. The effects of these changes on phenotypic and ultrastructural appearance, transcriptional profile and structure of chromatin is being analyzed.

72 Publications: Przewłoka M. and Jerzmanowski A. (1997) Improved method of total histone isolation from Arabidopsis thaliana. Biol. Plant. 39, 299-302. Prymakowska-BosakM, Przewłoka M., Iwkiewicz J., Egierszdorff S., Kuraś M., Chaubet N., Gigot C, Spiker S. and Jerzmanowski A. (1996) Histone HI overexpressed to high level in tobacco affects certain developmental programs but has limited effects on basal cellular functions. Proc. Nati. Acad. Sei. USA 93, 10250- 10255.

DEPARTMENT OF PLANT GROWTH AND DEVELOPMENT

Head: professor Andrzej Podstolski

1. METABOLIC CONTROL OF SEED DORMANCY AND GERMINATION

S. Lewak, R. Bogatek-Leszczyńska, B. Żarska-Maciejewska Dormancy in seeds is defined as a block (or a set of bocks) imposed on one (or several) cardinal process(es) of germination. Our research aim is to characterize these processes, to understand their regulation, and to explain the action of environmental factors affecting dormancy. Our attention is focused on the identification of control points of the pathways involved in the mobilization of seed reserves (storage lipids, proteins and sugars). Recently, the role of light, low temperature and cyanide in the control of triacylglycerol degradation (hydrolysis and b-oxidation) and transformations (gluconeogenesis) was studied in detail in apple embryos. The involvement of these factors in the control of sugar (native reserves and newly synthesized) catabolism in the same material was studied as well. In parallel to the studies on apple seeds, the moblization of storage lipids and proteins was investigated in sunflower seeds subjected to osmopriming. Publications: Corbineau F., Bogatek R., Radice S., Picard M-A and Come D., 1997, Changes in in vivo and in vitro ACC oxidase a ctivities during chilling and subsequent warming as exemplified by Vigna radiata seedlings. In: Biology and Biotechnology of the Plant Hormone Ethylene, Kanellis et al., eds, Kluwer Academic Publishers.

2. REGULATION OF SECONDARY METABOLITE BIOSYNTHESIS AND ACCUMULATION IN PLANTS

A. Podstolski, R. Dembowska, K. Heller

Biosynthetic pathway of vanillin and related C6-Ci metabolites in Vanilla planifolia tissue culture is studied. It was determined that biosynthesis of benzoate derivatives branches out of the phenylopropanoid pathway at the level of p-coumaric acid. Activity of p-coumarate chain shortening enzyme (p-hydroksybenzaldehyd

73 synthase) was found to be a key factor linking benzoate and phenylopropanoid pathways. The resulting /5-hydroxybenzaldehyde, due to high activity of aromatic alcohol dehydrogenase is reduced and stored as p-hydroxybenzyl alcohol. Our goal is to increase p-hydroxybenzaldehyde accumulation (by reducing aromatic alcohol dehydrogenase activity) and to stimulate the enzyme activities (polyphenol oxidases, monooxigenases) responsible for hydroxylation of p-hydroxybenzaldehyde at position 3, in order to shift Q - C\ metabolism towards immediate vanillin precursors. Publications: Havkin - Frenkel D., Podstolski A. and Knorr D. (1996). Effect of light on secondary product formation by cultures of Vanillaplanifolia . Plant Cell, Tissue and Organ Culture, 45, 133-136. Bronisz A. (1997). Factors influencing activity of aromatic alcohol dehydrogenase in cultured Vanilla planifolia tissues. M.Sci. thesis, University of Warsaw.

DEPARTMENT OF PLANT MORPHOGENESIS Head: professor Mieczyslaw Kuras

1. STRUCTURAL AND ULTRASTRUCTURAL INVESTIGATIONS OF ZYGOTIC AND SOMATIC PLANT MORPHOGENESIS

M. Kuras, R... Ilasz, A. Iwanowska, M. Stefanowska, T. Tykarska, A. Tylicki

Developmental regularities of zygotic and somatic embryogenesis and postembryogenesis of Brassica napns L. and Gentiana sp., organogenesis in in vitro culture of Solatium lycopersicoides; structural, ultrastructural and immunocytochemical investigations. Analysis and ultrastructural localization of secondary metabolites (mainly phenolic compounds); their correlation with activation processes and environmental stress influence. Publications: Zhang H.Q., Li Y.Q., Kuras M., Bednara J., Cresti M.. 1996. Influence of hydroxyurea on cell divisions and tubuline cytoskeleton in Allium cepa root meristem. Acta Soc. Bot. Pol. 65, 179-185. Prymakowska-Bosak M., Slusarczyk J., Przewtoka M, Jerzmanowski A., Kuras M. 1997. The anatomical and ultrastructuarl characterization of leaves of the transgenic tobacco overexpressing histone HI. Acta Biol. Sil. 3J_, 92-99 (in Polish) Tykarska T. 1997. Developmental regularities of zygotic and somatic embryos of Brassica napus L. Acta F. R. N. Univ. Comen.- Physiol. Plantarum. 3_9, 85-94.

74 ENVIRONMENTAL PLANT POLLUTION LABORATORY

Head: dr. Malgorzata Wierzbicka

THE INFLUENCE OF HEAVY METALS ON PLANTS

M. Wierzbicka, D. M. Antosiewicz, E. Michalak Plants accumulate large amounts of lead in their tissues without revealing any symptoms of poisoning. The question arises: what processes provide plants with such an effective protection against lead. Experiments were performed to check the effect of relative air humidity on the response of Allium cepa L. plants to lead. The differences in heavy metal tolerance of Allium cepa L. plants developing from bulbs and seeds were studied. The relationship between lead tolerance and mineral status of plants with special attention on the role of calcium metabolism was also investigated. Publications: Antosiewicz D.M., Schachtman D., Clemens S. and Schroeder J. 1996. Plant root cDNA enhances heavy metal uptake into yeast. Plant Physiol. 111 suppl: 54.

75 RESEARCH GRANTS

1. Research projects financed by Polish State Committee for Scientific Research (KBN)

1.1. Joanna Rytka "The role of ferrochelatase in the control of metabolism by heme. Isolation and characterization of Saccharomyces cerevisiae mutant strains with changed ferrochelatase activity"; 1993-1996

1.2 Kazimierz Kleczkowski "Potato transformation", 1993-1996;

1.3. Marta Dobrzanska "Attempts to construct an artificial chromosome for transformation of monocotyledonous plants", 1993-1997;

1.4. Tadeusz Klopotowski "Regulation of enterobacterial metabolism by temperature and its biophysical and medical aspects", 1993-1996;

1.5. Grazyna Muszynska "Characterization of plant kinase C and tyrosine protein kinases and involvement of these enzymes in signal transduction", 1994-1996;

1.6. Kazimierz Lech Wierzchowski "Structure-activity relationship of a non-regulated E.coli promoter in transcription initiation", 1994-1997;

1.7. Jaroslaw Kusmierek "Molecular basis of mutagenesis: study of the influence of neighboring bases in the template on coding properties of modified bases during DNA replication", 1994-1997;

1.8. Jacek Hennig "Salicylic acid-dependent cis- and trans-acting elements involved in the expression of PR genes", 1994-1997;

1.9. Monika Hryniewicz "New aspects of sulfur metabolism regulation in Enterobacteriaceae: relationship between cbl gene function and cysteine regulon", 1994-1997;

1.10. Danuta Hulanicka "Mechanisms of gene expression of potato leafroll virus", 1994-1996;

76 1.11. Maria Agnieszka Siwecka "Nuclease associated with rye germ ribosomes degrading double stranded RNA: purification, activators and inhibitors", 1994-1997;

1.12. Andrzej Paszewski "Sulphur amino acid metabolism in fungi: molecular characterization of structural and regulatory genes", 1994-1997;

1.13. Teresa Zohjdek "Mechanism of protein distribution to various cellular compartments in the yeast Saccharomyces cerevisiae", 1995-1997;

1.14. Wlodzimierz Zagorski "The structural and functional analysis of the genome of the yeast Saccharomyces cerevisiae", 1995-1998;

1.15. Magdalena Boguta "Nuclear control of mitochondrial function in yeast", 1995-1998;

1.16. Celina Janion "Modification of bases - induction of stable DNA replication", 1995-1998;

1.17. Andrzej Jerzmanowski "Studying the mechanism of Hl-mediated specific repression of the genes located in the vicinity of AT-rich DNA sequences", 1995-1997;

1.18. Iwona Fijalkowska "Fidelity of replication of the leading and the lagging DNA strands in E.coli", 1995-1998;

1.19. Zygmunt Ciesla "Isolation of a novel Saccharomyces cerevisiae gene, expression of which is induced by damage of DNA", 1995-1998;

1.20. Piotr Ceglowski "Gene functioning in the stability region segB from plasmid pSM19035", (1995-1998);

1.21. Witold Jachymczyk "The role of DNA polymerases in repair of various kinds of damage in DNA of Saccharomyces cerevisiae", 1995-1998;

1.22. Anna G6ra-Sochacka "Molecular biology of viroid infection - pathogen diagnostic and construction of resistsnt plants", 1995-1997;

77 1.23. Jan Brzeski ,,Molecular characteristics of Polish populations of Bursaphelenchus sp. nematodes living in pine wood", 1996-1997;

1.24. Jaroslaw Kusmierek "Mechanism of action and potential inhibitors of 3-methyladenine-DNA glycosylase of bacterial and human origin", 1996-1998;

1.25. Anna Przykorska "RNA structure probing with plant nucleases. Cloning of genes coding for Rn and ChS nucleases", 1996-1998;

1.26. Anna Kurlandzka ,,The search for the function of the essential S.cerevisiae gene U17918", 1996-1998;

1.27. Jacek Bardowski ,,Cellobiose - induced P-galactosidase activity in Lactococcus laclis", 1996-1998;

1.28. Wlodzimierz Zagorski "Constructions of virus-resistant potato transformants", 1996-1998;

1.29. Anna Chachulska "Construction of transgenic plants resistant to potato virus Y", 1996-1998;

1.30. Danuta Hulanicka ,,Analysis of transgene expression in potato lines transformed with potato leafroll virus cDNA", 1996-1997;

1.31. Maria Bucholc ,,Isolation, recombination and application of linear plasmids in plant transformation", 1996-1998;

1.32. Michat Dadlez "Rational design and production of specific serine proteinase inhibitors based on molecular modeling, kinetic data analysis and site directed mutagenesis", 1996-1998;

1.33. Tadeusz Chojnacki "Rubber - like polyprenoid lipids of plant origin", 1996-1998;

78 1.34. Andrzej Ozyhar (Wroclaw Politechnic) Krystyna Grzelak "Expression of protein elements of functional ecdysteroid receptor in bacterial and yeast cell", 1996-1999;

1.35. Tadeusz Kulikowski "Synthesis, investigation of the structure and conformation of new antitumor, antiviral, and antiparasitic nucleosides and nucleotides and invesigation of their interactions with thymidylate synthase", 1996-1999;

1.36. Agnieszka Halas "The role of DNA polymerases in gene conversion in the yeast S.cerevisiae", 1996;

1.37. Grazyna Palamarczyk "Improvement of protein production-secretion in Trichoderma reesei", 1997-1999;

1.38. Agnieszka Chaciiiska "Interaction of yeast mitochondrial ribosomal protein Nam9 with chaperone HsplO4", 1996;

1.39. Joanna Michalik ,,The nuclear polyhedrosis virus from Stilpnotia salicis (Lymantriidar) - more effective biopesticide. Genome characteristics and virulence", 1997-2000;

1.40. EwaBartnik "The molecular basis of mitochondrial diseases", 1997-1999;

1.41. Agnieszka Sirko "Oral vaccines from plants: construction of transgenic plants producing urease of Helicobacterpylori", 1997-1999;

1.42. Iwona Smaczynska-de Rooij "Yeast S.cerevisiae mutants defective in the regulation of the peroxisomal proteins expression - isolation and characterization", 1997;

1.43. Irena Pietrzykowska "SOS dependent mutagenesis: possible mechanism of down regulation of the SOS response and role of UmuD'C proteins in mutagenesis independent of DNA replication", 1997-1999;

1.44. Ewa Kula-Swiezewska "Prenylation of plant proteins", 1997-1999;

79 1.45. Urszula Maciejewska "Somatic hybridization - pathogen resistance in potato", 1997-2000;

1.46. Anna Szkopiriska "Genetic and biochemical characterization of polyprenol synthesis in eukaryotes", 1997-2000;

1.47. JacekHennig "Study of virus resistance in transgenic potato plants expressing replicase gene sequence of potato leafroll virus", 1997-2000;

1.48. Ryszard Otlewski (Wroclaw University) Michal Dadlez "Selection from protein libraries on M13 phage applied for production of new proteinase inhibitors and optimization of their structural specifity and folding", 1997-2000;

1.49. rLobocka Malgorzata ,,Studies on partition of low copy number plasmids to daughter cells and cell division", 1997-1999;

1.50. Joanna Rytka ,,The hem mutants of the yeast S.cerevisiae as a tool in studies of the interacellular transport and regulatory role of heme", 1996-1998;

1.51. Andrzej Jerzmanowski ,,Studies on the function of histone HI in eukaryotic cell, the identification of protein factors responsible for transcriptional accessibility of plant chromatin and the analysis of the mechanism of SWI/SNF dependent transcription", 1997-2000;

1.52. Grazyna Dobrowolska ,,The role of protein kinases in transduction of salinity stress in plants", 1997-1999;

2. International co-financed Research Projects

2.1. Research projects KBN-French Embassy

2.1.1. Danuta Hulanicka "Reverse genetics of potato leafroll luteovirus (PLRV)", 1995-1996;

2.1.2. Joanna Michalik "Construction of recombinant baculovirus as an insecticide", 1995-1996; 2.1.3. Grzegorz Przewlocki "Posttranscriptional regulation of CFTP gene expresion in human colon carcinoma cell line and novel sequence specific RNase", 1996-1997;

2.1.4. Grazyna Palamarczyk ,,Genetic and biochemical characterization of dolichol chain synthesis in eucaryotic cells", 1996-1999;

2.1.5. Jacek Bardowski ,,Genetic diversity of bacteria from Lactococcus genus" , 1996-1997;

2.2. Research projects KBN-The British Council

2.2.1. Grazyna Palamarczyk ,,Characterization of glycosylation-deficient mutants of Aspergillus nidulans", 1994-1996;

2.2.2. Andrzej Paszewski "Molecular biology of sulphur metabolism in Aspergillus nidulans", 1995-1997;

2.2.3. Marta Dobrzanska "The development and evaluation of plant artificial chromosomes", 1995-1997.

3. Research projects financed by International Organizations

3.1. NATO Grant Joanna Michalik Jadwiga Chroboczek (ISB, CEDEX 1) "Introduction of the baculovirus protein expression system to Poland", 1994-1996;

3.2. NATO Grant Hilary Koprowski (USA) Grazyna Muszynska Agnieszka Sirko ,,Construction of transgenic plants producing immunogenic proteins of therapeutic significance", 1996-1997;

3.3. Project Polish-German Cooperation Fund Wieslaw Jankowski "Isolation and practical application of long-chain polyisoprenoids", 1994-1997;

81 3.4. Project COPERNICUS Pia Ek (Uppsala University) Grazyna Muszyriska ,,Metabolic regulation in plants. Specificity of signal transduction by the phospholipid-dependent protein kinase from seedlings and prediction of potential substrates of this enzyme among plant proteins", 1997-1999;

3.5. Project COPERNICUS A. Leone (RCVBNC - Italy) Alina Kacperska (Warsaw Uniwersity) Grazyna Muszyriska ,,Manipulation of the lipid composition and physical state of plant membranes for improving tolerance to temperature stress", 1997-1999;

3.6. Project Maria Sktodovvska-Curie Found II Iwona Fijalkowska ,,Mutagenesis in Leading and Leading Strands of DNA Replication", 1996-1998;

3.7. Polish-Israeli project financed by UNESCO Piotr Zielenkiewicz ..International Center for Cooperation in Bioinformatics", 1996-1997

3.8. Project PHARE SCI-TECH Wlodzimierz Zagorski Andrzej Jerzmanowski (Warsaw University) ,,Cooperation between Institute of Biochemistry and Biophysics PAS and the Faculty of Biology, Warsaw University in teaching of molecular biotechnology courses", 1996-1997;

3.9. Project Stefan Batory Fundation Piotr Zielenkiewicz Michal Minczuk (student, Warsaw University) ,,Molecular biology in the Internet", 1996-1997;

82 MEETINGS, SYMPOSIA, COURSES (organized by Institute of Biochemistry and Biophysics)

1996

,,Gene Structures and Expression in Plant Pathogenes" prof. S. Lewak, 18-22 January, Warszawa

,,Two Faces of Oxygen" prof. A. Podstolski, 26 April, Warszawa

,,Polish-French Course on Bioinformatics" dr. P. Zielenkiewicz, 3-5 June, Warszawa

Workshop on ,,Genes and their Products in Basic Research and Biotechnology" (Third Survey of Research), 28-30 November, Warszawa

1997

,,Diversity and Unity in Biological Sciences" (International TEMPUS Course), 5-17 May, Warszawa

,,Mechanisms of DNA Repair and Mutagenesis" (On the 100-th Anniversary of the Discovery of Polonium and Radium) dr. B. Tudek, 8-11 October, Warszawa

,,lCCBnet Bioinformatics Training Workshop for Central and Eastern Europe" (Under the auspices of UNESCO and the Polish State Committee for Scientific Research) dr. P. Zielenkiewicz, 27-30 October, Warszawa

,,Molecular Basis of Signal Transduction in Plants" prof. S. Lewak, 28-29 November, Warszawa

,,Molecular Mechanisms of Insect Ontogenesis Regulation" (Dedicated to the memory of prof. Z. Lassota), dr. K. Grzelak, 16 December, Warszawa

83 INSTITUTE SEMINARS Presented by Staff Members of IBB PAS and (*) Invited Guests.

1996

1. Synthesis and secretion in yeast of the human epidermal growth factor (huEGF). J. Topczewska; 09.01.96

2. Expression of heterologous proteins and peptides in the baculovirus system. J. Michalik; 16.01.96

3.* Saccharomyces cerevisiae, an economical genome. Ch. Herbert / Centre de Genetique Moleculaire, Gif-sur-Yvette, France/; 16.01.96

4.* NKAM- a new gene in killer cells. M. Kozlowski / University of Arkansas, Medical Sciences, USA/; 26.01.96

5. The gene coding for TBP (TATA Binding Protein) in Aspergillus nidulcms. R. Kucharski; 30.01.96

6. Overproduction of histone HI from A. thaliana in tobacco plant. M. Prymakowska - Bosak; 13.02.96

7. Maturation of mRNA precursors. Genetic analysis and manipulations with antisense oligonucleotides. Z. Domiriski; 20.02.96

8. Modeling of intramolecular electron transfer in model peptides. J. Poznanski; 27.02.96

9.* Progress in the molecular analysis of the rye B-chromosome. T. Langdon /Institute of Biological Sciences, The University of Wales, Aberystwyth, U.K.; 05.03.96

10.* DNA base excision repair. T. O' Connor /Institut Gustave Roussy, Villejuif, France/; 12.03.96

11.* Synthesis of linear and circular forms of human immunodeficiency virus proviral DNA. A. Pokrovski / Institute of Molecular Biology, Novosibirsk, Russia/; 14.03.96

12. Early stages in the folding of bovine pancreatic trypsin inhibitor. M. Dadlez; 02.04.96

84 13. Specific protein-protein interactions within Escherichia coli mutasome. A. Nowicka and P. Jonczyk; 16.04.96

14. The adenoviral penton. J. Chroboczek / Institut de Biologie Structurale, Grenoble, France/; 19.04.96

15. Analysis of streptokinase gene expression in bacterial homologous and heterologous systems. I. Kern; 23.04.96

16.* A search for new antitumor drugs based on a cytotoxity profile against selected cell-lines. P. Lassota / American Cyanamid Company, Pearl River, NY, USA/; 24.04.96

17. Site-directed mutagenesis of the HEM 15 gene encoding ferrochelatase in the yeast Sacharomyces cerevisiae. Physiological and biochemical characterization of mutants. M. Gora; 07.05.96

18. Synthesis of tioderivatives of fluorodeoxyuridines potential antitumour compounds. M. Bretner; 14.05.96

19.* Mechanisms of mutagenesis in E. coli. R. Schaaper /N1EHS, Research Triangle Park, USA/; 15.05.96

20. Molecular mechanism of the ecdysteroid action. K. Grzelak; 21.05.96

21.* Regulation of calcium dependent protein kinases. B. Kemp /St. Vincent's Institute of Medical Research, Australia/; 27.05.96

22. Photoreactions of psolarens with the cell membrane lipids: photoproduct formation during PUVA therapy. Z.Zanjbska; 28.05.96

23. A new gene of the cysteine regulon in Escherichia coli - cbl. R. Iwanicka; 11.06.96

24.* DNA etheno adducts as critical exposure markers and as markers of genotoxic effects associated with oxidative stress/lipid peroxidation. A. Barbin /International Agency for Research on Cancer, France/; 17.06.96

25. Magnesium induced assymetry of DNA opening in promoter complexes with bent helix axis. J. Smagowicz; 18.06.96

85 26.* The use of molecular biology in reference diagnosis of emerging infectious diseases. N. Pieniazek / Parasitic Diseases Branch, CDC, Atlanta, USA/; 19.06.96

27.* Co-evolution of host-parasite systems: a theory and selected examples. R. Korona / Jagiellonian University, Krakow/; 14.06.96

28.* Biochemistry and molecular biology of cold adaptation in plants: antifreeze proteins and mechanisms of gene targeting. D. Guerra /University of Idaho, Moscow, USA/; 24.06.96

29. Cloning and characterization of Aspergillus nidulans sulphur metabolism regulatory genes. R. Natorff; 25.06.96

30. Genetic variability in populations of microorganisms revealed by the Multilocus Enzyme Electrophoresis (MLEE)" K. Lewicka; 26.06.96

31.* Transcription proteins - mechanisms of specific interactions with DNA. P. Czernik /University of Arkansas, USA/; 01.07.96

32.* Nuclear matrix attachment regions (MARs) and their function in transgene expression in plant cells. S. Spiker/Dept. of Genetics, N. Carolina University, USA/; 25.07.96

33.* Antitumor ribonucleases. W. Ardelt /ALFACELL Corporation Bloomfield, NJ, USA/; 2.08.96

34.* Isolation and analysis of RNA. P. Chomczynski /Molecular Research Center, Inc., Cincinnati, USA/; 8.08.96

35.* A simplifed cell-free system to study the mechanism of pre-mRNA splicing. M. Konarska /The Rockefeller University, New York, USA/; 3.09.96

36.* Prion diseases. P. Liberski /Medical Academy, Lodz/; 24.09.96

37.* Emerging infectious diseases. J. Jeljaszewicz/National Institute of Higiene, Warsaw/; 30.09.96

38.* Do chromosomes need genes? N. Jones /Cell Genetics Laboratory, The University of Wales, UK/; 09.10.96

39.* Strategies for engineering plant viruses resistance. E. Balazs /Agricultural Biotechnology Center, Godollo, Hungary/; 10.10.96

86 40. Methods of protein structure prediction. P. Zielenkiewicz; 29.10.96

41. Before life had appeared. T. KJopotowski; 5.11.96

42. Plant ethylene receptor. J. Hennig; 12.11.96

43. New perspectives in studies on partition of low copy number plasmids at cell division. M.Lobocka; 3.12.96

44.* Retroviral integrase-crystal structure of a new target for designing drugs against AIDS. A. Wlodawer /NCI-FCRDC, Frederick, USA/; 16.12.96

45. Synthesis and a role of ethylene in plant responses to the stress factors. A. Kacperska; 17.12.96

1997

1. Protein structure prediction using threading. K. Pawfowski; 7.01.97

2. Chromatin structure and transcription. A. Jerzmanowski; 14.01.97

3. Recognition of tRNAs by aminoacyl tRNA - synthetases. A. Przykorska; 21.01.97

4. Approaches to making vaccines in plants. P. McGarvey/Thomas Jefferson University, Philadelphia, USA/; 28.01.97

5. The genome of PI bacteriophage. M.Lobocka; 28.01.97

6. Rearrangements of chromosome DNA of the yeast S. cerevisiae. R. Gromadka; 04.03.97

7.* DNA mismatch repair in E.coli. M. Marinus /University ofMassachuetts Medical School, Worcester, USA/; 10.03.97

8. Genetic regulation of folate metabolism in Aspergillus nidulans. I. Lewandowska; 11.03.97

87 9. DNA damage induced by free radicals - its influence on DNA replication and mutagenesis" M. Graziewicz; 18.03.97

10. * Mechanisms of nickel carcinogenesis. A. R. Oiler/NIPERA, Durham, USA/; 21.03.97

11. N15 bacteriophage of Escherichia coli - an evolutionary puzzle. M.Lobocka; 25.03.97

12. sconC - a regulatory gene of sulphur metabolism in Aspergillux nidulans. M. Piotrowska; 08.04.97

13.* Red pepper viruses in Tunisia. M. Marrakchi /Tunis University, (Faculty of Sciences, Tunisia)/; 10.04.97

14.* Quantitative modelling of biomolecular hydration. D. Mario Soumpasis /The Max-Planck Institute for Biophysical Chemistry, Gottingen/; 22.04.97

15. Fidelity of leading and lagging DNA strands synthesis in E.coli. I. FijaJkowska; 29.04.97

16. Calcium-phospholipid dependent protein kinases from maize seedlings. J. Szczegielniak; 20.05.97

17.* New in vivo models in oncology. P. Lassota /American Cyanamid Company, Pearl River, N.Y, USA/; 21.05.97

18.* Gene transfer by electroporation and liposomes. S.W. Hui /Roswell Park Cancer Institute, USA/; 23.05.97

19. From purple bacteria to man: ancient genes regulate RNA metabolism. P. Ste.pieti; 27.05.97

20. Mechanisms of stable maintenance of the pSM 19035 plasmid in bacterial cells. U.Zielenkiewicz; 10.06.97

21. Protein tyrosine kinases in plants. J. Trojanek; 17.06.97

22.* Adaptive responce to ionizing radiation. A. Wojcik /Department of Radiobiology, Institute of Nuclear Chemistry and Technology, Warsaw/; 16.06.97 23.* Fidelity studies with E. coli replication proteins. R. Schaaper /NIEHS, Research Triangle Park, USA/; 16.06.97

24. arcA - a regulator of arginine metabolism in Aspergillus nidulans. J. Empel; 24.06.97

25.* Recent approaches to cure type I diabetes. A. Sledziewski /Biopharmaceuticals - NovoNordisk, Seattle, USA/; 27.06.97

26.* Molecular analysis of the foldosome from higher plant cytoplasm. J. A. Miernyk /Mycotoxin Research Unit, Peoria II, USA/; 7.08. 97

27.* Plant viruses as tools for production of biomedicals in plants. V. Yosibov/Thomas Jefferson University, Philadelphia,USA/; 13.08.97

28.* The multifunctional role of coat protein in the replication of alfalfa mosaic virus. J. Bol /Institute of Molecular Plant Sciences, Leiden University, Holandia/; 30.09.97

29.* Studies on the synthesis and biological activities of isonucleosides. Li-He Zhang /Beijing Medical University, Beijing, ChRL/; 26. 09.97

30.* Towards automated protein crystallization diagnostics. Y. Georgalis /Institut fur Kristallographie, Freie Universitat, Berlin/; 16.10.97

31. From n-D NMR towards 3-D protein structure. A. Ejchart; 21.10.97

32. Why should one study the biology of IncP plasmids ? G. Jagura-Burdzy; 28.10.97

33.* Prediction of bendability and curvature in genomic DNA. S. Pongor/ICGEB, Trieste, Italy/; 29.10.97

34. Mechanisms of protein crystallization. A. Kierzek; 04.11.97

35. Pencillin binding proteins of Streptococcus pneumoniae. I. Kern; 18.11.97

36. Higher plant homologues of the bacterial chaperones. B. Kroczyriska; 02.12.97

37. Rye germ ribosome - associated nuclease II: substrate specificity and structure. M. Siwecka; 09.12.97

89 38.* Role of E. coli cellular proteases in elimination of proteins aggregated in vivo by heat shock A. Taylor / Chair of Biochemistry, University of Gdansk, Gdansk/; 09.12.97

39.* Deciphering of biological functions of protein phosphatases 2A by analysis of copurifying proteins and application of specific inhibitors" S. Zolnierowicz /Biotechnology Faculty, School of Medicine and University of Gdansk, Gdansk/; 15.12.97

40. Structure and function of plant protein phosphatase type 2A (PP2A). A. O. Sunday; 16.12.97

90 LIST OF PUBLICATIONS

This list comprises publications of Staff Members of the Institute in 1996-1997. Publication numbers are referred to in reports, above. Publications resulting from individual external collaborations of staff members are marked + (e.g. +3141); abstracts of oral and poster presentations at scientific meetings are marked A (e.g. 890/A) and patents are marked P (e.g. P4). The list includes also titles of Dissertations for M. Sc, Ph. D. and Doctor Habilitatus (Dr. Sc.) degrees. M. Sc. degrees are issued by collaborating Institutions of Higher Education, Ph. D. and Dr. Sc. - by the Institute. PUBLICATIONS

3099. TROJANER J., EK P., SCOBLE J., MUSZYŃSKA G., ENGSTRÖM L. Phosphorylation of plant proteins and the identification of protein tyrosine inase activity in maize seedlings. Eur. J. Biochem. (1996) 235, 338-344

3 102. HECHT K., ZHANG S., KŁOPOTOWSKI T., AMES G. F. L. D-Histidine utilization in Salmonella typhimurium is controlled by the eucine-responsive regulatory protein (Lrp). J. Bacterio!. (1996)J78, 327-331

3103. BĘBENEK A., PIETRZYKOWSKA I. The isfA mutation inhibits mutator activity and processing of UmuD protein in Escherichia coli recA730 strains. Mol. Gen. Genet. (1996) J250, 674-680

3105. BOBROWSKI K., SCHÖNEICH CH. Decarboxylation mechanism of the N-terminal glutamyl moiety in y-glutamic acid and methionine containing peptides. Radiât. Phys. Chem. (1996) 47, 507-510

3106. GRZESIUK E., JANION C. MMS-induced mutagenesis and DNA repair in Escherichia coli dnaQ49: Contribution of UmuD' to DNA repair. Mutation Res. (1996) 362, 147-154

3 109. PAWLOWSKÍ P., SZUTOWICZ I., RÓŻYCKI S., ZIELIŃSKI J., FIKUS M. Bioelectrorheological model of the cell. VI. Experimental verification of the rheological model of cytoplasmic membrane. Biophys. J. (1996) 70, 1024-1026

3110. TOPCZEWSKA J., REMPOŁA B., FIKUS M. Expression of small synthetic genes coding for hEGF, human epidermal growth factor, and CPTI II, serine proteinase inhibitor from Cucurbitacea, cloned in a novel expression/secretion vector in Saccharomyces cerevisiae. Acta Biochim. Polon. (1996) 43, 255-264

3111. ŁOZIŃSKI T., WIERZCHOWSKI K.L.

Effect of reversed orientation and length of An • Tn DNA bending sequences in the -35 and spacer domains of a consensus-like Escherichia coli promoter on its strength in vivo and gross structure of the open complex in vitro. Acta Biochim. Polon. (1996)43,265-280

92 3112. DZIK J. M., BRETNER M., KULIKOWSKI T., GOŁOS B., JARMUŁA A., POZNAŃSKI J., RODE W., SHUGAR D. Synthesis and interactions with thymidylate synthase of 2,4-dithio analogues of UMP and 5-fluoro-dUMP. Biochim. Biophys. Acta (1966) 1293, 1-8

3113. STOLARSKI R., SITEK A., STĘPIŃSKI J., JANKOWSKA M., OKSMAN P., TEMERIUSZ A., DARŻYNKIEWICZ E, LÖNNBERG H., SHUGAR D. ^H-NMR studies on association of mRNA cap-analogues with tryptophan- containing peptides. Biochim. Biophys. Acta (1996) 1293, 97-105

3114. FELCZAK K., DRABIKOWSKA A. K. VILPO J. A., KULIKOWSKI T., SHUGAR D. 6-substituted and 5,6-disubstituted derivatives of undine: stereoselective synthesis, interaction with uridine phosphorylase, and in vitro antitumor activity. J. Med. Chem. (1996) 39, 1720-1728

3115. HENNIGJ. Elements of signal leading to activation of plant defense (in Polish). Biotechnologia (1996)32,40-51 3116. WĘGRZYN A., WĘGRZYN G., TAYLOR K. Disassembly of the coliphage X replication complex due to heat shock induction of the groE operon. Virology (1996) 2T7, 594-597

3118. SHUGAR D. The NTP phosphate donor in kinase reactions: Is ATP a monopolist? Acta Biochim. Polon. (1996) 43, 9-24

3119. CAFFIERI S., ZARĘBSKA Z., DALL 'ACQUA F. Psoralen photosensitization: damages to nucleic acid and membrane lipid components. Ibid.: 241-246

3120. DOMIŃSKI Z., KOLE R. Effects of exon sequences on splicing of model pre-mRNA substrates in vitro Ibid.: 161-174

3121. BUKOWSKA A. M., KUŚMIEREK J. T. Miscoding properties of isoguanine (2-oxoadenine) studied in an AMV reverse transcriptase in vitro system. Ibid.: 247-254

93 3123. JOŃCZYK P, NOWICKA A., Specific in vivo protein-protein interactions between Escherichia coli SOS mutagenesis proteins. J. Bacteriol. (1996)178 2580-2585

3124. PAWŁOWSKI K., BIERZYŃSKI A., GODZIK A. Structural diversity in a family of homologous proteins. J. Mol. Biol. (1996)258 349-366

3125. GALIBERT F., ALEXANDRAKI D., BAUR A, BOLES E., TOVAN D., CHALWATZIS N., CHUAT J.C., COSTER F., CZ1EPLUCH C, DE HAAN M., DOMDEY H., DURAND P., ENTIAN K.D., GATIUS M., GOFFEAU A., GRIVELL L.A., HENNEMANN A., HERBERT C.J., HEUMANN K., HILGER F., HOLLENBERG, C.P., HUANG M.E., JACQ C, JAUNIAUX i.C, KATSOULOU C, KIRCHRATH L., KLEINE K., KORDES E., KÖTTER P., LIEBL S., LOUIS E.J..MANUS V, MEWES H.W., MIOSGA T., OBERMAIER B, PEREA J., POHL T., ROSSAU R., PORTETELLE D., PUJOL A., PURNELLE B., RAMEZANI RAD M., RASMUSSEN S.W. ROSE M., SCHAFF-GERSTENSCHLÄGER I., SMITS P.H.M., SCARCEZ T., SORIANO N., TZERMIA M., VAN BROEKHOVEN A., VANDENBOL M., WEDLER H., VON WETTSTEIN D., WAMBUTT R., ZAGULSKI M., ZÖLLNER and KARPFINGER-HARTL L. Complete nucleotide sequence of Saccharomyces cerevisiae chromosome X. EMBOJ. (1996)15,2031-2049 3126. GÓRA M., GRZYBOWSKA E., RYTKA J, LABBE-BOIS R. Probing the active-site residues in Saccharomyces cerevisiae ferrochelatase by directed mutagenesis. J. Biol. Chem. (1996)271, 11810-11816

3127. SKOCZYŁAŚ E., ŚWIEŻEWSKA E., Protein farnesyltransferase in plants. Biochimie (1996) 78, 139-143

3128. GROMADKA R., GÓRA M., ZIELENKIEWICZ U„ SŁONIMSKI P. P., RYTKA J. III Yeast Sequencing Reports. Subtelomeric duplications in Saccharomyces cerevisiae chromosomes III and XI:Topology, arrangements, corrections of sequence and strain-specific polymorphism. Yeast (1996) 12, 583-591

3129. SKONECZNY M., RYTKA J. Maintenance of the peroxisomal compartment in glucose-repressed and anaerobically grown Saccharomyces cerevisiae cells. Biochimie (1996) 78, 95-102

94 3130. GÓRA A., CHACINSKA A., RYTKA J., LABBE-BOIS R. Isolation and functional characterization of mutant ferrochelatases in Saccharomyces cerevisiae. Ibid: 144-152

3131. WÓJCIK A., GRZESIUK E., TUDEK B., JANION C. Conformation of plasmid DNA from Escherichia coli deficient in the repair systems protecting DNA from 8-oxyguanine lesions. Ibid.: 85-89

3132. SZKOPIŃSKA A., KARST F, PALAMARCZYK G. Products of S. cerevisie c/s-prenyltransferase activity in vitro. Ibid.: 111-116

3133. JANKOWSKI J. M., CANNON P. D. van der HOORN F, WASILEWSKA L.D., WONG N. C.W., DIXON G. H. Regulation of protamine gene expression in an in vitro homologous system. Acta Biochim. Polon. (1996) 43, 369-378

3134. KRUSZEWSKA J. S., PERLIŃSKA-LENART U., PALAMARCZYK G. Solubilization and one-step purification of mannosylphosphodolichol synthase from Trichoderma reesei. Acta Biochim. Polon. (1996) 43, 397-402

3135. GROMADKA R., KANIAK A., SŁONIMSKI P. P., RYTKA J. A novel cross-phylum family of proteins comprises a KRR1 (YCLO59c) gene which is essential for viability of Saccharomyces cerevisiae cells. Gene (1996) 171,27-32

3136. KAMIEŃSKA-TRELA K., WÓJCIK J. Applications of spin-spin couplings (invited review). Nucl. Magn. Reson. (1996) 25, 177-228

3137. WIERZCHOWSKI J., WIELGUS-KUTROWSKA B., SHUGAR D. Fluorescence emission properties o 8-azapurines and their nucleosides, and application to the kinetics of the reverse synthetic reaction of purine nucleoside phosphorylase. Biochim. Biophys. Acta (1996) 1290, 9-17

3138. BZOWSKA A., KULIKOWSKA E., POOPEIKON.E., SHUGAR D. Kinetics of phosphorolysis of 3- (/?-D-ribofuranosyl) adeninę and 3-(/MD- ribofuranosyl) hypoxanthine, non-conventional substrates of purine- nucleoside phosphorylase. Eur. J. Biochem. (1996) 239, 229-234

95 3139. SZYSZKA R., BOGUSZEWSKA A., SHUGAR D., GRANKOWSKI N. Halogenated benzimidazole inhibitors of phosphorylation, in vitro and in vivo, of the surface acidic proteins of the yeast ribosomal 60S subunit by endogenous protein kinases CK-II and PK60S. Acta Biochim. Polon. (1996) 43, 389-396

3140. PASZEWSKI A., BALIŃSKA M. Regulation of folate metabolism in fungi: Aspergillus nidulans as an experimental model (in Polish). Post. Biochem. (1996) 42, 129-132

+3141. FIJAŁKOWSKA 1. J.,SCHAAPER R. M. Mutants in the Exo I motif of Escherichia coli dna Q: Defective proofreading and inviability due to error catastrophe. Proc. Nati. Acad. Sei. USA (1996) 93, 2856-2861

3 142. POOPEIKO N. E., DRAB1KOWSKA A. K.,POZNAŃSK1 JL SHUGAR D, KULIKOWSKI T. Synthesis, conformation and biological properties of 2\ 3'-dideoxy- 3'- fluoro-5-chloro-4-thiouridine, potential anti-HIV agent. 10 "' Symposium on the Chemistry of Nucleic Acid Components. September 1st-7st, 1996 Treat' Castle Czech Republic. Collect. Czech. Chem. Commun. (1996) 6J_, S16-S19

3143. ŚLEDZIEWSKA-GÓJSKA E. Repair of O"-methyIguanine-DNA (in Polish). Post. Biochem. (1996) 42,21-30

3144. SADOWY E., ZAGÓRSKI W., HULANICKA D. Infectious cDNA clones of plant viruses (in Polish). Biotechnologia (1996) 34, 60-68

3 145. STANKIEWICZ M, REMPOŁA B., FIKUS M. 3 ' noncoding sequences of the CTA 1 gene enhance expression of the recombinant serine protease inhibitor, CPTIII, in Saccharomyces cerevisiae. Acta Biochim. Polon. (1996) 43, 525-530

3146. PRYMAKOWSKA-BOSAK M, PRZEWLOKĄ M. R., IWKIEWICZ J., EGIERSZDORFF S., KURAŚ M., CHAUBET N., GIGOT C, SPIKER S., JERZMANOWSKI A. Histone HI overexpressed to high level in tobacco affects certain developmental programs but has limited effect on basal cellular functions. Proc. Nati. Acad. Sei. USA (1996) 93, 10250-10255

96 3147. ONO B. I., KIJIMA K, ISHII N., KAWATO T., MATSUDA A., PASZEWSKI A., SHINODA S. Regulation of sulphate assimilation in Saccharomyces cerevisiae. Yeast (1996) Y2, 1153-1162

3148. LEWANDOWSKA I., BALIŃSKA M., NATORFF R., PASZEWSKI A. Regulation of folate-dependent enzyme levels in Aspergillus nidulans: studies with regulatory mutants. Biochim. Biophys Acta (1996) 1290, 89-94

3149. BURZYŃSKA B., TOPCZEWSKI J. Genotyping of Bison bonasiis /c-casein gene following DNA sequence amplification. Animal Genetics (1996) 26, 335-336

3150. LAPIŃSKI L., PRUSINOWSKA D., NOWAK M. J., BRETNER M., FELCZAK K., MAES G., ADAMOWICZ M. Infrared spectra of 6-azathiouracils: an experimental matrix isolation and theoretical ab initio SCF/ 6-311 G study. Spectrochimica Acta Part A (1996) 52, 645-659

3151. CALIKOWSKI T. T. Chromatin complexes in regulation of gene expression during development (in Polish). Post. Biochem. (1996) 42, 238-243

3152. BOGUTA M. Nuclear control of mitochondrial functions in Saccharomyces cerevisiae (in Polish). Post. Biochem ( 1996) 42, 259-268

3153. PŁOCHOCKA D., WEŁNICKI M., ZIELENKIEWICZ P., ZAGÓRSKI-OSTOJA W. Three-dimensional model of the potyviral genome-linked protein. Proc. Nati. Acad. Sei. USA (1996) 93, 12150-12154

+3154. MALAMY J, SÁCHEZ-CASAS P, HENNIG J., GUO A., KLESSIG D. F. Dissection of the salicylic acid signaling pathway in tobacco. Mol. Plant-Microbe Interact. (1996) 9, 474-482

3155. BRODZIK R, HENNIG J. Effect of DNA methylation on transient expression of PR-s genes in tobacco protoplasts. Genet. Pol. (1996)37A, 105-109

97 3 156. HUG G. L., MARCINIAK B., BOBROWSKI K. Acid-base equilibria involved in secondary reactions folloving the 4-carboxybenzophenone sensitized photooxidation of methionylglycine in aqueous solution.Spectral and time resolution of the decaying (S.#. N)+ radical cation. J. Phys.Chem. (1996)100, 14914-14921

3157. WOLUCKA B. A., ROZENBERG R., de HOFFMAN E., CHOJNACKI T. Desorption chemical jonization tandem mass spectrometry of polyprenol and dolichyl phosphates. J. Am. Soc. Mass Spectrom. (1996) 7, 958-964

3 158. KROCZYŃSKA B., ZHOU R., WOOD C, M1ERNYK J. A., AtJl, a mitochondrial homologue of the Escherichia coli DnaJ protein. Plant Mol. Biol. (1996) 31, 6^19-629

3 159. GÓRA A., CANDRESSE T., ZAGÓRSKI W. Use of intramolecular chimeras to map molecular determinants of symptom severity of potato spindle tuber viroid (PSTVd). Arch. Virol. (1996) HI, 2045-2055

+3 160. LI D„ DOBRO WOLSKA G., KREBS E. G. The physical association of caséine kinase 2 with nucleolin . J. Biol. Chem. (1996) 27J_, 15662-15668

+3161. MEGG1OF., DEANA A. D., RUZZENE M., BRUNATI A. M., CESARO L, GUERRA B., MAYER T., METT H., FABBRO D., FURET P., DOBROWOLSKA G., PINNA L. A. Different susceptibility of protein kinases to staurosporine inhibition. Kinetic studies and molecular bases for the resistance of protein kinase CK2. Eur. J. Biochem. (1995) 234, 317-322

3162. PRZYKORSKAA. RNA structure probing with use of two plant nucleases (in Polish). Ed: Fundacja. Rozwój SGGW. Warszawa (1996) p. 59

3163. CHACHULSKA A. M., ZAGÓRSKI W. Sequence analysis of genomes of potato virus Y isolates (in Polish). In:"Selected problems of plant biotechnology" Materials for biotechnology students, edited by K.Wypijewski .Lectures given in frame of Open Seminars of the Institute of Molecular Biology and Biotechnology AMU, Poznan, 16 February-1 June (1995)

98 3164. BUCHOWICZJ. Plant minichromosomes (in Polish). Ibid: 149-158

3165. KUCHARSKI R., BARTNIK E. TBP - structure, functions and interactions with other transcription factors (in Polish). Post. Biol. Komorki (1996) 23, 67-80.

3166. WE.GRZYN A., TAYLOR K, W^GRZYN G. The cbpA chaperone gene function compensates for dnaJm X plasmid replication during amino acid starvation of Escherichia coli. J. Bacteriol. (1996) 178, 5847-5849

3167. WE.GRZY7M G., WE.GRZYN A., PANKIEWICZ A., TAYLOR K. Allele specificity of the Escherichia coli dnaA gene function in the replication of plasmids derived from phage X. Mol. Gen. Genet. (1996) 252, 580-586

3168. SZALEWSKA-PALASZ A., WE.GRZYN A., OBUCHOWSK1 ML, PAWLOWSKI R. B1ELAWSKI K., THOMAS M.S., WE.GRZYN G. Drastically decreased transcription from CH-activated promoters is responsible for impaired lysogenization of the Escherichia coli rpoA341 mutant by bacteriophage X. FEMS Microbiol. Lett. (1996) 144,21-27

+3169 WEICHSEL A., MONTFORD W. R., CIESLA J., MALEY F. Promotion of purine nucleotide binding to thymidylate synthase by a potent folate analogue inhibitor, 1843U89 Proc. Natl. Acad. Sci. USA (1995) 92, 3493 - 3497

+3170. DADLEZ M., KIM P. S. Rapid formation of the native 14-38 disulfide bond in the early stages of BPTI folding. Biochemistry (1996) 35, 16153-16164

3171. BARDOWSKIJ. Molecular biology in healthy food production (in Polish). Przemysl Spozywczy (1996) 50, 41-43

3172. THIRD SURVEY OF RESEARCH. Abstracts of the Workshop on: Genes and their products in basic research and biotechnology. Institute of Biochemistry and Biophysics PAS, Warszawa, 28-30 November 1996

99 3173. WĘGLENSKA A., JACOB B., SIRKO A. Transcriptional pattern of Escherichia coli ihfB {him D) gene expression. Gene (1996) HI, 85-88

+3174. GRĄZIEWICZ M., WINK D. A., LAVAL F. Nitric oxide inhibits DNA ligase activity : potential mechanisms for NO-mediated DNA damage. Carcinogenesis (1996)17^ 2501 -2505

3175. DRABIKOWSKA A. K. Uridine phosphorylase from Hymenolepis diminuta (Cestoda): Kinetics and inhibition by pyrimidine nucleoside analogs. Acta Biochim. Polon. (1996)43, 733-742

3176. KULCITSKY V., HERTEL J., SKOCZYŁAŚ E., ŚWIEŻEWSKA E., CHOJNACKI T. The occurence of long-chain polyprenols in leaves of plants of Combretaceae family. Ibid.: 707-712

3177. ŚWIEŻEWSKA E., CHOJNACKI T. Polyprenols in leaves of fruit-trees of Rosaceae family. Ibid.: 701 -706

3178. KŁUDKIEWICZ B., GODLEWSKI J., GRZELAK K., CYMBOROWSKI B. LASSOTA Z. Influence of low temperature on the synthesis of some Galleria mellonella proteins. Ibid.: 639 - 644

3179. TOMASZEWSKI R, JERZMANOWSKI A. The AT-rich flanks of the oocyte-type 5 S RNA gene of Xenopus laevis act as a strong local signal for histone HI- mediated chromatin reorganization in vitro. Nucleic Acids Res.(1997) 25,458 - 465

3180. GAVRYUSHOV S., ZIELENKIEWICZ P. Multivalent ion distribution around a cylindrical polyion: Contribution of polarization effects due to difference between dielectric properties of the macro molecule's interior and the agueous solvent. J. Phys. Chem. B (1997) jjn, 792 - 797

100 3181. GORA-SOCHACKA A., KIERZEK A., CANDRESSE Th., ZAGÓRSKI W. The genetic stability of potato spindle tuber viroid (PSTVd) molecular variants. RNA (1997) 3, 68-74

3182. WIELGUS-KUTROWSKA B., KULIKOWSKA E., WIERZCHOWSKI J., BZOWSKA A., SHUGAR D. Nicotinamide riboside, an unusual, non typical, substrate of purified purine - nucleoside phosphorylases. Eur. J. Biochem. (1997) 243, 408 - 414

3183. ŚWIEŻEWSKA E., SZYMAŃSKA M., SKORUPIŃSKA K., CHOJN ACKI T. The occurrence of long chain polyprenols in leaves of plants. In: Physiology, Biochemistry and Molecular Biology of Plant Lipids Eds: J. P. Williams, M. U. Khan and N. W. Lern. Kluwer Acad.Publishers. Toronto 1996 pp.192-194

3184. WÓJCIK J.,GÓRAL J., PAWŁOWSKI K., BIERZYŃSKI A. Isolated calcium-binding loops of EF-hand proteins can dimerize to form a native-ike structure. Biochemistry (1997) 36, 680-687

3185. POKORSKI M., WALSKI M., MATYSIAK Z. A phospholipase C inhibitor impedes the hypoxic ventilatory response in the cat. In: Frontiers in Arterial Chemoreception. Eds, P. Zapata, C. Eyzaguirre, and W. Torrance . Adv. Exp. Med. Biol. (1996) 410, 397-403

3186. STROKOVSKAYA L., ZIEMNICKA J., MICHALIK J. Genetic variability of four natural isolates of the Stilpnotia salicis multiple - enveloped nuclear polyhedrosis virus. Acta Biochim. Polon. (1996) 43, 633-638

3187. MICHALIK J., CHOJNICKA B., CYMBOROWSKI B Vitellogenesis in virgin and mated females of the mealworm beetle, Tenebrio molitor. Ibid.: 623-632

3188. MIECZKOWSKI P. A., FIKUS M. U., CIEŚLA Z. Characterization of a novel DNA damage-inducible gene of Saccharomyces cerevisiae, DIN 7, which is a structural homolog of the RAD2 and RAD27 DNA repair genes. Mol. Gen. Genet. (1997) 253, 655-665

101 3189. SLEDZIEWSKA-GÓJSKA E., TORZEWSKA D. Different repair of O6-methylguanine occurring in DNA modified by MMS in vivo or in vitro. Mutation Res. (1997)381, 31-37

3190. DADLEZM. Hydrophobie interactions accelerate early stages of the folding of BPT I. Biochemistry (1997) 36, 2788-2797

3191. BIRNBAUM K. B., KIERDASZUK B., SHUGAR D. Cristal structure of l,3,5-trimethyl-N4-hydroxy-cytosine, and its relevance to the mechanism of hydroxylamine mutagenesis. Nucleosides Nucleotides ( 1996) 15, 1805-1819

3192. SA WERS G., KAISER M., SIRKO A., FREUNDLICH M. Transcriptional activation by FNR and CRP: reciprocity of binding-site recognition. Mol. Microbiol. (1997) 23, 835-845

3193. ŻOŁĄDEK T., TOBIASZ A., VADUVA G., BOGUTA M., MARTIN N. C, HOPPËR A. K. MDP1, a Saccharomyces cerevisiae gene involved in mitochondrial / cytoplasmic protein distribution, is identical to the ubiquitin-protein ligase gene RSP5. Genetics (1997) 145, 595-603

3194. SIRKO A., WĘGLEŃSKA A., HRYNIEWICZ M., HULANICKA D. M. Characterization of the Escherichia coli gene encoding a new member of short-chain dehydrogenase / reducíase (SDR) family. Acta Biochim Polon. (1997) 44, 153-158

3195. BOGUTA M., CZERSKA K., ŻOŁĄDEK T. Mutation in a new gene MAFI affects tRNA suppressor efficiency in Saccharomyces cerevisiae. Gene (1997) 181, 291-296

3196. JUSZCZUK M., ZAGÓRSKI-OSTOJA W., HULANICKA D. M. Studies on the translation mechanism of subgenomic RNA of potato leafroll virus. Acta Biochim. Polon. (1997) 44, 69-78

3197. BUCHOWICZJ. Nuclear extrachromosomal DNA of higher plants. Ibid.: 13-20

102 3198. SZURMAK B, BUCHOWICZ J. Autonomous replication of a wheat DNA sequence in isolated wheat nuclei. Ibid.: 79-82

3199. PRZE WLOK A M. R., JERZMANOWSKI A. Improved method of total histone isolation from Arabidopsis thaliana. Biol. Plant. (1997) 39, 299-302

3200. Editorial Professor David Shugar awarded Honory Doctorate from the University of Warsaw. Post. Biochem. (1997) 43, 2-4

3201. WOJCIK A, JANION C. Mutation induction and mutation frequency decline in halogen light- irradiated Eschericha coli K-12 AB1157 strains. Mutation Res. (1997) 390, 85-92

3202. SIWECKA M. A. Purification and some properties of a novel dsRNA degrading nuclease bound to rye germ ribosomes. Acta Biochim.Polon. (1997) 44, 61-68

3203. KIERDASZUK B, MODRAK-WOJCIK A, SHUGAR D. Binding of phosphate and sulfate anions by purine nucleoside phosphorylase from Escherichia coli :ligand-dependent quenching of enzyme intrinsic fluorescence. Biophys. Chem. (1997) 63_, 107-118

3204. KUCHARSKI R., BARTNIK E. The TBP gene from Aspergillus nidulans - structure and expression in Saccharomyces cerevisiae. Microbiology (1997) 143, 1263-1270

3205. IWANCZAK F, PYTRUS T., KASZUBSKA-POLKOWSKA 1, TREMBACZ H. Loesch-Nyhan syndrom in a 5-year-old boy (in Polish). Ped. Pol. (1997) 72, 183-185

3206. HALAS A., BARANOWSKA H., POLICINSKA Z., JACHYMCZYK W. Involvement of the REV3 gene in the methylated base-excision repair system Co-operation of two DNA polymerases, 8 and Rev3p, in the repair of MMS- induced lesions in the DNA of Saccharomyces cerevisiae. Curr. Genet. (1997) 3J_, 292-301

103 3207. BOBROWSKI K, HOLCMAN J, POZNANSKI J., WIERZCHOWSKI K.L. Pulse radiolysis studies of intramolecular electron transfer in model peptides and proteins . 7.Trp°-»TyrO° radical transformation in hen egg-white lysozyme. Effects of pH, temperature, Trp62 oxidation and inhibitor binding. Biophys. Chem. (1997)63, 153-166

3208. CHACHULSKA A. M, CHRZANOWSKA M., ROBAGLIA C, ZAGORSKI W. Tobacco veinal necrosis determinants are unlikely to be located within the 5' and 3'terminal sequences of the potato virus Y genome. Arch.Virol. (1997) J42, 765-779

3209. DOBRZANSKA M, KRYSIAK C, KRASZEWSKA E. Transient and stable transformation of wheat with DNA preparations delivered by a biolistic method. Acta Physiol. Plant. (1997) I£, 277-284

3210. SHUGARD. Biophysics Department of the University of Warsaw and development of molecularbiology (in Polish). Post. Fizyki (1997) 48, 139-143

3211. TOBIASZ A., ZOLADEK T. The ubiquitin conjugation system and ist role in the yeast Saccharomyces cerevisiae (in Polish). Post. Bioch. (1997) 43, 91-97

3212. KAMIENSKA-TRELA K., WOJCIK J. Applications of spin-spin couplings. (Invited review) Nucl. Magn. Reson. (1997) 26, 173-226

3213. KRZYMOWSKA M., HENNIG J. Simple and rapid technique to detect PVY presence in some Solanaceae plants. Acta Physiol. Plant. (1997) 19, 95-99

3214. PAWLOWSKI K., JAROSZEWSKI L., BIERZYNSKI A., GODZIK A. Multiple model approach - dealing with alignment ambiguities in protein modeling In: Pacific Symp.on Biocomputing 1997. Eds: R. B.Altman, A. K. Dunker, L. Hunter T. E. Klein. World Scientific pp.328-339

3215. BOLEWSKA K., KOZLOWSKA H., GOCH G., MIKOLAJEK B., BIERZYNSKI A. Molecular cloning and expression in Escherichia coli of a gene coding for bovine S100A1 protein and its Glu32->Gln and Glu73-»Gln mutants. ActaBiochim. Polon. (1997) 44, 275-284

104 3216. PLESNARZ., MALANOWSKI S., ŁOTOWSKI Z., MORZYCKIJ. W., FRELEK1, WÓJCIK J. Stereospecific association of C-20 epimers of 3 /?-hydroxy-16-oxo-24-nor-17- azachol-5-eno-23-nitryle. Z. Naturforsch. (1997) 52b, 749-756

3217. SZKOP1ŃSKA A, GRABIŃSKA K., DELOURME D., KARST F., RYTKA J., PALAMARCZYK G. Polyprenol formation in the yeast Saccharomyces cerevisiae : effect of farnesyl diphosphate synthase overexpression. J. Lipid Res. (1997) 38, 962-968

3218. KIERZEK A. M, WOLF W. M., ZIELENK1EWICZ P. Stimulations of nucleation and early growth stages of protein crystals. Biophys. J. (1997)73, 571-580

3219. ŻOŁĄDEK T., NGUYEN B1CH NH1, JAGIEŁŁO 1., GRACZYK A., RYTKA J. Diamino acid derivatives of porphyrins penetrate into yeast cells, induce photodamage, but have no mutagenic effect. Photochem. Photobiol. (1997) 66, 253-259

3220. AW AN M. F. M., MAC1EJEWSKA U., KLECZKOWSKI K., WIELGAT B. Differential responses of potato cell suspensions to a culture filtrate of Phytophthora infestans. Z. Naturforsch. (1997) 52c, 333-338

3221. FIJAŁKOWSKA 1. J., JOŃCZYK P. Coordination of the leading and lagging DNA strand synthesis (in Polish). Post. Biochem. (1997) 43, 98-104

3222 DANIK1EW1CZ W., OLEJNIK M, WÓJCIK J., TYRLIK S. K, NALEWAJKO B. Methyleneglucoses - transition metal catalyzed synthesis from formaline and glucose; importance of heterobimetallic catalyst. J. Mol. Cat. A:Chem. (1997) 123, 25-33

3223. BARDOWSKI J., LEWICKA K. Modern methods of identification and differentiation of dairy starters (in Polish). Przegląd Mleczarski (1997) 9, 300-302

105 3224. GORA-SOCHACKA A., PAWŁOWICZ J., CANDRESSE Th., ZAGÓRSKI W. Structure of the pathogenicity domain and disease symptoms induced by potato spindle tuber viroid (PSTVd). Bull. Pol. Ac. Biol. (1996) 44, 273-277

3225. DZIK J. M, ZIEL1ŃSKI Z., JARMUŁA A., MICHALSKI R., RODE W., LES A., BRETNER M., FELCZAK K., KULIKOWSKI T. Interactions of thymidylate synthase with 5-hydroxy-dUMP and 5-hydroxymethyl-dUMP, and their 4-thio analogues. In: Chemistry and Biology of Pteridines and Folates 1997, Proel 1th Intern. Symp. on Pteridines and Folates, Berchtesgaden, Germany, June 15-20,1997, 415-418 Eds: Wolfgang Pfleiderer, Hartmut Rokos

3226. GOŁOS B., DZIK J. M., RODE W., JANKOWSKA J., KRASZEWSKI A., STAWIŃSKI J, SHUGAR D. Interaction of thymidylate synthase with the 5'-thiophosphates and 5'-H- phosphonates of 2'-deoxyuridine, thymidine and 5-fluoro-2'-deoxyuridine. Ibid.: 423-426

3227. ŻOŁĄDEK T., NGUYEN BICH NHI, RYTKA J. Saccharomyces cerevisiae mutants defective in heme biosynthesis as a tool for studying the mechanism of phototoxicity of porphyrins. Photochem.Photobiol. (1996) 64, 957-962

3228. CHACHULSKA A. M., FAKHFAKH H, ROBAGLIA Ch., GRANIER F., ZAGÓRSKI W., VILAINE F. Synthesis of full-lenght poty virus cDNA copies suitable for the analysis of genome polymorphism. J.Virol. Methods (1997) 67, 189-197

3229. MICHALIK J. Baculoviruses - alternative to chemical pesticides (in Polish). Pestycydy (1996) 3, 23-30

3230. JANION C. Dynamic mutations (in Polish). Kosmos (1997) 46, 213-220

3231. SYLLER J., MARCZEWSKI W., PAWŁOWICZ J. Transmission by aphids of potato spindle tuber viroid encapsidated by potato leafroll luteovirus particles. Eur. J. Plant Patol. (1997) 103, 285-289

106 3232. LEWANDOWSKA I., BALIŃSKA M., PASZEWSKI A. Identification of new regulatory genes controlling synthesis of folate- dependent enzymes in Aspergillus nidulans. Microbiology (1997) 143. 3273-3278

3233. TOPCZEWSKI J., SIEŃKO M, PASZEWSKI A. Cloning and characterization of the Aspergillus nidulans cysB gene encoding cysteine synthase. Curr. Genet. (1997) 31., 348-356

+3234. LI D., MEIER U. Th., DOBROWOLSKA G., KREBS E. G. Specific interaction between casein kinase 2 and the núcleo lar protein Noppl40. J. Biol. Chem. (1997)272, 3773-3779

3235. BOGUTA M., CHACIŃSKA A., MURAWSKI M., SZCZEŚNIAK B. Expression of the yeast NAM9 gene coding for mitochondrial ribosomal protein. ActaBiochim Polon. (1997) 44 , 251-258

3236. BRETNER M, FELCZAK K., DZIK J. M., GOŁOS B., RODE W., DRABIKOWSKA A., POZNAŃSKI J., KRAWIEC K., PIASEK A., SHUGAR D., KULIKOWSK1 T. Thiated pyrimidine deoxynucleoside analogues, potential chemotherapeutic agents, and substrates / inhibitors in various enzyme systems. Nucleosides Nucleotides (1997) 16, 1295-1299

3237. DADLEZ M. Disulflde bonds in protein folding studies :friends or foes? Acta Biochim. Polon. (1997) 44, 433-452

3238. PARMRYD I., SHIPTON C. A., ŚWIEŻEWSKA E., DALLNER G., ANDERSSON B Chloroplastic prenylated proteins. FEBS Lett. (1997) 4J4, 527-531

3239. SINDELAR P. J., CHOJNACKI T., VALTERSSON C. Role of apolipoprotein A-IV in hepatic lipase-catalyzed dolichol acylation and phospholipid hydrolysis. Biochemistry (1997) 36, 1807-1813

3240. PAWŁOWSKI P., POZNAŃSKI A., FIKUS M. Bioelectrorheological model of the cell. 7.Cellular deformation in the presence of cytochalasin B. Biorheology(1997)34, 171-193

107 +3241. FIJAŁKOWSKA I. J., DUNN R. L., SCHAAPER R. M. Genetic requirements and mutational specificity of the Escherichia coli SOS mutator activity. J. Bacteriol. (1997)23, 7435-7445

3242. WALIŃSKA K, JANAS T., CHOJNACKI T, ŚWIEŻEWSKA E., JANAS T. Properties of lipid bilayers modified by long-chain polyprenols. Cell. Mol. Biol. Lett. (1997) 2, 89-100

3243. BIERZYŃSKI A., PAWŁOWSKI K Helix-coi! transition theories. Are they correct? Acta Biochim. Polon. (1997) 44, 423-432

3244. FIKUS M. Methods of DNA recombination in vitro (in Polish). In: Biotechnologia Zwierząt p. 18-59 PWN 1997

3245. TOCZYŁOWSKA B., ZGUKOV I., DĘBICKI G. High resolution 'H NMR spectroscopy of human cerebrospinal-fluid: a preliminary study. Med. Sei. Monit. (1997) 3, 404-409

3246. KIERZEK A., ZIELENKIEWICZ P. Energy minimization of globular proteins with solvent effects included. Comparison of empirical solvation energy terms and explicit water treatment. Acta Biochim. Polon. (1997) 44_, 549-556

3247. OBUCHOWSKI M, WĘGRZYN A., SZALEWSKA -PAŁASZ A., THOMAS M. S., WĘGRZYN G., An RNA polymerase a subunit mutant impairs N-dependent transcriptional antitermination in Escherichia coli. Mol. Microbiol. (1997) 23, 211-222

3248. OBUCHOWSKI M., GILADI H., KOBY S., SZALEWSKA -PAŁASZ A., WĘGRZYN A., OPPENHEIM A. B.,THOMAS M. S., WĘGRZYN G. Impaired lysogenisation of the Escherichia coli rpoA341 mutant by bacteriophage X is due to the inability of CII to act as a transcriptional activator. Mol. Gen. Genet. (1997) 254, 304-311

3249. WĘGRZYN G., WĘGRZYN A. Activation of transcription in Escherichia coli (in Polish). Post. Biol. Kom. (1997) 24 (Supl.8), 53-68

108 3250. CHACHULSKA A. M., CHRZANOWSKA M, FLIS B., KRZYMOWSKA M., LIPSKA-DWUZNIK A, ROBAGLIA Ch., ZAGORSKI W. Potato and tobacco cultivars transformation towards potato virus Y resistance (in Polish). Biotechnologia(1997) 4, 48-54

3251. CHRZANOWSKA M, DOROSZEWSKA T., CHACHULSKA A. Characteristics of PVY strains occuring in Poland on tobacco and potato crops (in Polish). Prog. Plant Protect./Poste_py w Ochronie Roslin (1997) 37, 326-328

3252. PALUCHA A., CHRZANOWSKA M, ZAGORSKI W., HULANICKA D. Genetically engineered potato resistant to potato leafroll virus infection (in Polish). Biotechnologia (1997) 4, 38-47

3253. BOBROWSKI K., POZNANSKI J., WIERZCHOWSKI K. L. Long range electron transfer between proline-bridged aromatic amino acids. In: Photochemistry and Radiation Chemistry. Eds. D. G. Nocera, J. F. Wishart, American Chemical Society. 131-143

3254. WIERZCHOWSKI K.L. Intramolecular electron transfer between tryptophan radical and tyrosine in oligoproline-bridged model peptides and egg-white lysozyme. Acta Biochim. Polon. (1997) 44, 627- 644

3255. WE_GRZYN A., WE_GRZYN G., HERMAN A., TAYLOR K. Protein inheritance:^, plasmid replication perpetuated by the heritable replication complex. Genes to Cells (1996) I, 953-963

3256. TAYLOR K., WE_GRZYN G., WE.GRZYN A, SZALEWSKA- PALASZ A., HERMAN A., OBUCHOWSKI M., SRUTKOWSKA S., KONOPA G. Escherichia coli initiator protein DnaA, cell-cycle and control of X plasmid replication. Mini review. Bull. Pol. Ac. Biol. (1996) 44, 225-230

3257. GEORGALIS Y., UMBACH P, ZIELENKIEWICZ A, UTZIG E, ZIELENKIEWICZ W., ZIELENKIEWICZ P., SAENGER W. Microcalorimetric and small-angle light scattering studies on nucleating lysozyme solutions. J. Am. Chem. Soc. (1997) JJ_9, 11959-11965

109 3258. ZATYKA M, JAGURA-BURDZY G, THOMAS Ch. M. Transcriptional and translational control of the genes for the mating pair formation apparatus of promiscuous IncP plasmids. J. Bacteriol. (1997) 179., 7201-7209

3259. JAGURA-BURDZY G., THOMAS Ch. M. Dissection of the switch between genes for replication and transfer of promiscuous plasmid RK2: Basis of the dominance oftrfAp over trbAp and specificity for KorA in controlling the switch. J.Biol.Mol. (1997)265., 507-518

3260. Van der PLOEG J. R., IWAN1CKA-NOWICKA R., KERTESZ M. A., LEISINGER T, HRYNIEWICZ M. M. Involvement of CysB and Cbl regulatory proteins in expression of the tauABCD operon and other sulfate starvation -inducible genes in Escherichia coll J. Bacteriol. (1997)179, 7671-7678

3261. SHUGAR D., BOROWSKI E. Eds. of Symp. Molecular Aspects of Chemotherapy. Proc. of 6th Intern. Symp. on Mol. Aspects of Chemotherapy, Gdansk, Poland, July 9-12, 1997 Pharmacol. Ther. (1997) 76,1-242 Special Issue

3262. GAVRYUSHOV S., ZIELENKIEWICZ P. Electrostatic potential and free energy of proteins: A comparison of the Poisson-Boltzmann and the Bogolyubov-Born-Green-Yvon equations. J. Phys. Chem. B (1997) KM, 10903- 10909

3263. BARTN1KE. The human mitochondrial genome - mutations, polymorphisms and diseases (in Polish). Post. Biol. Kom. (1997) 24, supl. 69-75

3264. KNOESTER M., HENNIG J, van LOON L.C., BOL J. F., LINTHORST H. J. M. Isolation and characterization of tobacco cDNA encoding an ETR1 homolog. (Accession No AF022727) Plant Physiol. (1997)111, 1731- 1733

3265. OTLEWSKI J., APOSTOLUK W., BUCZEK O., CHOLAWSKA L., GRZESIAK A., KOSCIELSKA K, KROKOSZYNSKA 1, KROWARSCH D., STACHOWIAK D., DADLEZ M. Structural and energetic aspects of protein inhibitor-serine proteinase recognition.

110 In: Trends in Protein Research. Polish - Japanese Seminar Poznan, 18-19 November, 1997 eds: Andrzej B.Legocki and Kenji Soda Sei. Publisher OWN Poznan 1997 49-58

3266. BIERZYŃSKI A. cc-helix formation as an early step of protein folding: A critical examination of the helix-coil transition theories. Ibid.: 107-110

3267. PRYMAKOWSKA-BOSAK M., TOMASZEWSKI R., PRZEWLOKĄ M., ŚLUSARCZYK J., DŁUŻNIEWSKA J., CHAUBET N., GIGOT C, SPIKER S., KURAŚ M., JERZMANOWSKI A. Histone HI - a well known protein with the unknown function. Ibid.: 27-30

3268. LINDBLOM S., EK P., MUSZYŃSKA G., EK B., SZCZEGIELNIAK J., ENGSTRÖM L. Phosphorylation of sucrose synthase from maize seedlings. Acta Biochim. Polon. (1997) 44. 809-818

3270. HULANICKA D, ZAGÓRSKI-OSTOJA W., PALUCHA A. Non-convential strategies to protect plants against viral infections (in Polish). Zeszyty Naukowe Akademii Rolniczej 1997, Kraków

3271. KAZIMIERCZAK B., HERTEL J., ŚWIEŻEWSKA E., CHOJNACKI T., MARCZEWSKI A. On the specific pattern of long chain polyprenols in green needles of Pinus mugo Turra. Acta Biocbim. Polon. (1997) 44, 803-808

+ 3272. RIEGER K. J., KANIAK A., COPPEE J. Y., ALJINOVIC G., BAUDIN- BAILLIEU A., ORŁOWSKA G., GROMADKA R., GROUDINSKY O., DI RAGO J. P, SŁONIMSKI P. P. Yeast Functional Analysis Reports. Large-scale phenotypic analysis - the pilot project on yeast chromosome III. Yeast (1997) 13, 1547-1562

111 ABSTRACTS. MISCELLANEOUS.

890/A MIKOLAJCZYK M., SZCZEGIELNIAK J., GAGANIDZE D., JURKOWSKI I., DOBROWOLSKA G., MUSZYNSKA G. Protein kinases involved in stress signalling in plants. Intern. Summer School on "Molecular Mechanisms of Signaling and Targeting". Island of Spetsai, Greece, 18-30 August 1996 post. Abstr. p.42

89I/A BARDOWSKI J. Lactic acid bacteria - from basic research to modern applications (in Polish). Europ. Conf. under auspices of the program ,,Biotechnology" 22-26 October 1995 Cork, Ireland; Przegla_d Mleczarski, January 1, 1996, 24-25

892/A KRZYMOWSKA M. Free radicals in plant pathogenesis (in Polish). Conference:"Two faces of oxygen", organized by Polish Botanical Society, Warsaw University and IBB PAS. 26.04.1996

893/A BARDOWSKI J. Modern methods of identification of lactic acid bacteria (in Polish). Summer School:"Lactic Acid Bacteria: classification, metabolism, genetics, applications" Dobieszkow, 19-23 May 1996 program W.2

894/A BARDOWSKI J. Basics of bacterial genetics (in Polish). Ibid.: program W.7

895/A BARDOWSKI J. The use of recombinant DNA technology to improve lactic acid bacteria (in Polish). Ibid.: program W.ll

896/A BARDOWSKI J. Characterization of the lactose operon from lactic acid bacteria and its potential applications (in Polish). Ibid.: program W.9

112 897/A FRANK S, CAFFIERI S, VEDALDI D., ZARĘBSKA Z., DALL'ACQUA F. Structure activity relationship between psoralen-oleic acid photocycloadducts and PKC activity. Photochem. Photobiol. (1996) 63, Special Issue 24th Annual Meeting June 15-20, 1996 Atlanta, Georgia Abstr. MPM-H1 p.55S

898/A RAYA R. R, BARDOWSKI J., EHRLICH S. D., CHOPIN A. Multiple transcriptional control of the tryptophan biosynthetic operon of Lactococcus lactis. 5 Symposium on Lactic Acid Bacteria-Genetics, Metabolism and Applications. FEMS, Veldhoven, The "Netherlands, 8-12 September 1996 Abstr. H 28

899/A ALEKSANDRZAK T., BARDOWSKI J. Cellobiose-induced b-galactosidase activity in Lactococcus lactis IL 1403 Ibid.: H 36

900/A CZERNIEĆ E., BARDOWSKI J. Plasmid-encoded starch-degrading enzyme in Lactococcus lactis strain of non dairy origin. Ibid.: H 37

901/A BARDOWSKI J. Mechanisms of regulation of gene expression in Lactococcus lactis (in Polish). XXIIIld Meeting of Polish Microbiological Society, Łódź 1996 Abstr. p.76

902/A ALEKSANDRZAK T., CZERNIEĆ E, BARDOWSKI J. Localization of genes encoding cryptic ß-galactosidase and amylase in Lactococcus lactis cells (in Polish). Ibid.: P-006 p.84

903/A BIERZYŃSKI A. Three-dimentional structure of globular proteins (in Polish). XXXIInd Meeting of Polish Biochemical Society, 17-20 September, 1996 Krakow Abstr. IV-1 p.l 10, Plenary lecture

904/A NATORFF R., PIOTROWSKA M., PASZEWSKI A. SCONB, an Aspergillus nidulans sulphur metabolism regulatory gene encodes a b-transducin-like protein. ECFG3 3 European Conference on Fungal Genetics Münster, Germany, March 27-30, 1996 p.64 nr 43:B

113 905/A GRZESIUK E., JANION C. MMS-induced mutagenesis and DNA repair in E.coli defective in mismatch repair. DNA Repair Network Workshop. Smolenice Castle, Slovakia 29 April-1 May, 1996 p. 15, L 5

906/A WOJCIK A., JANION C. Halogen light induced mutagenesis and mutation decline in E.coli AB 1157 strains. Ibid.: p.46, P 12

907/A MACIEJEWSKA U., AWAN M. F. M. Oxidative response to Phytophthora infestans elicitor in Solanum tuberosum and S. nigrum suspension cells. 10 FESPP Congress "From Molecular Mechanisms to the Plant: An Integrated Approach". Florence, Italy 9-13 September 1996 p.285 S19-12

908/A ZARE_BSKA Z., WASZKOWSKA E., POZNANSKI J. Photoadducts of psoralen to lecithins: their possible role in the photochemotherapy PUVA. 12th International Congress on Photobiology (ICP'96) September 1-6 1996 Hofburg Congress Center Vienna, Austria abstr. p.206, 038

909/A PIETRZYKOWSKA I., CZAJKOWSKA A. A possible mechanism of UV-induced mutagenesis of 1 sus 08 in the nonpermissive for phage DNA replication host. Ibid: p.216, 080

910/A PIETRZYKOWSKA I., BE.BENEK A., CZAJKOWSKA A. Mechanisms of UV induced mutagenesis in Escherichia coli. "DNA Repair Network Workshop" Cancer Res. Instit. Slovak Academy of Sci. Smolenice Castle, Slovakia 29 April-1 May, 1996 abstr. L7 p.17

91 I/A PIETRZYKOWSKA I. Mechanism of DNA repair - organisms defence against environmental mutagens (in Polish). XXXIInd Meeting of Polish Biochemical Society, 17-20 September, 1996 Krakow Abstr. II-2 p.31, Plenary lecture

912/A BE_BENEK A., PIETRZYKOWSKA I. Influence of isfA mutation on UV-induced transitions in recAr and transversions in recA730 strains (in Polish). Ibid.: abstr. H-7 p.35

114 913/A HETMNIG J., BRODZIK R. Methylation of deoxyadenosine affects transient expression from the tobacco PR-1A promoter. 8th Intern.Congress "Molecular Plant-Microbe Interactions". Knoxville, TN July 14-19, 1996. Abstr. M-12

914/A CHACHULSKA A. M, ROBAGLIA C, DINANT S., ROSA I, ZAGORSKI W. Construction of full-length cDNA copies of potato virus Y (PVY) isolates. XXXIInd Meeting of Polish Biochemical Society, 17-20 September, 1996 Krakow. Abstr. 1-18

915/A OBUCHOWSKI M, SZALEWSKA-PALASZ A, WE_GRZYN A. A role for the a subunit of RNA polimerase in transcriptional antitermination (in Polish). Ibid.:IV-25, p. 124

916/A WE_GRZYN A., TAYLOR K., WE_GRZYN G. The cbpA gene function in X phage and plasmid DNA replication (in Polish). Ibid.: IV-20,p.l22

917/A WE_GRZYN G, WE_GRZYN A., PANKIEWICZ A., TAYLOR K. Allele specificity of dnaA in X plasmid replication (in Polish). Ibid.: IV-19, p.121

918/A WE.GRZYN A., WE_GRZYN G., TAYLOR K. The mechanisms of the disassembly of bacteriophage X replication complex upon heat shock (in Polish). Ibid.: IV-18, p. 121

919/A WE.GLENSKA A., SIRKO A., HULANICKA D. Characterization of the new gen ucpA localised on 52 'of E. coli chromosome (in Polish). Ibid.: XI-102, p.310

920/A KLUDKIEWICZ B., GRZELAK K. Influence of 20-hydroxyecdysone on protein sythesis in Galleria mellonella larvae (in Polish). Ibid.: XI-101, p.310

92I/A GORA A., PAWLOWICZ J., ROSA I, ZAGORSKI W. Phenotypically different potato spindle tuber viroid molecular variants vary in the secondary structure of the P domain. Ibid.: IV-43, p. 133

115 922/A PAWLOWICZ J. M, GORA A, ROSA I., CHACHULSKA A. M, WELNICKI M. Construction of plasmids for plant transformation in the aim of getting resistance to PSTVd infections (in Polish). Ibid.: IV-42, p.133

923/A GRAZIEWICZ M. A., ZASTAWNY T. H., OLINSKI R., JANION C, TUDEK B. DNA replication on templates modified with hydroxyl radicals generating system hypoxantine-xantine oxidase (in Polish). Ibid.: VIII-7, p. 209

924/A TREMBACZ H., JEZEWSKA M. M. The variety of adenine nucleoside phosphorylases (in Polish). Ibid.: XI-13, p. 266

925/A SIWECKA M. A., TOMASZEWSKI M., NAPIERALA M., KRZYZOSIAK W. J. Specificity of ribosomal nuclease of rye germ (in Polish). Ibid.: XI-123, p.321

926/A WE.GRZYN A., WE^GRZYN G., TAYLOR K. Is RNA polymerase p subunit a transcriptional activator contact site? (in Polish). Ibid.:IV-28,p.l26

927/A SZCZEGIELNIAK J., JURKOWSKI1., DOBROWOLSKA G., MUSZYNSKA G. Calcium and phospholipid dependent protein kinase from maize seedlings (in Polish). Ibid.: VI-19, p.171

928/A KAZIMIERCZAK B, MARCZEWSKI A., SWIEZEWSKA E, CHOJNACKI T. The influence of morphological and anatomical differences of dwarf mountain pine (Pinus mugo, Turra) on the polyprenol pattern (in Polish). Ibid.: VI-18, p. 171

929/A SZKOPINSKA A., GRABINSKA K., PALAMARCZYK G. Mutation of yeast FPP synthase encoding gene results in accumulation of unusually long chain polyprenols (in Polish). Ibid.: VI-16,p.l70

116 930/A GRABOWSKA D., SKONECZNY M., SZKOPINSKA A. Presence of peroxysomal structures is not a prerequisite for polyprenol synthesis in Saccharomyces cerevisiae (in Polish). Ibid.: VI-17, p. 170

93 l/A SZOŁAJSKA E, MICHALIK J. Baculovirus recombinant as insecticid (in Polish). Ibid.: II-63, p. 63

932/A HOFFMAN-ZACHARSKA D., PAPWORTH M., STĘPIEŃ P. Cloning and analysis of gene for glucoamylase of Aspergillus awamori (in Polish). Ibid.: 1-17, p. 13

933/A PRZEWŁOCKI G., EDELMAN A., PRZYKORSKA A. New RNase from colonie carcinoma cell line. Ibid.: III-20, p.81

934/A LENART U., LIWOS A., PALAMARCZYK G. Invertase secretion in Aspergillus nidnlans strains impaired in mannosylphosphodolichol synthase (in Polish). Ibid.: VI-10, p. 167

935/A KRUSZEWSKA J., PALAMARCZYK G. Modulation of Trichoderma reesei pathway by overexpression of yeast mannosylphosphodolichol synthase gene (in Polish). Ibid.: VI-2, p. 159

936/A MUSZYŃSKA G. Phosphorylation of plant proteins (in Polish). Ibid.: VI-8, p.165 Plenary lecture.

937/A IWANICKA-NOWICKA R., HRYNIEWICZ M.M., HULANICKA M.D. The function of Cbl protein in the expression of cys PTWAM operon of E. coli (in Polish). Ibid.: XI-107, p.313

938/A ŚWIEŻEWSKA E., SZYMAŃSKA M., SKORUPIŃSKA K., CHOJNACKI T. The occurrence of long chain polyprenols in leaves of plants. 12th Intern.Symp.on Plant Lipids. July 7-12, 1996 Toronto Canada. Abstr. B49

117 939/A SWIEZEWSKA E., CHOJNACKI T. The occurrence of long chain polyprenols in leaves of plants. Intern.Symp. on Isoprenoid Biochemistry. Zao, Japan, August 20 - 24, 1996 Plenary lecture.

940/A SWIEZEWSKA E., SZYMANSKA M., SKORUPINSKA K., HERTEL J., KULCITSKY V, CHOJNACKI T. The occurrence of the longest chain polyprenols in leaves of plants. 37th Intern.Conf. on the Biochemistry of Lipids . Antwerp (Belgium) September 4-7, 1996 Abstr. P222 p.89

941/A TROJANEK J., MUSIELAK M., MUSZYNSKA G. Tyrosine phosphorylation in proteins of maize seedlings. 15th Annual Symp. Current Topics in Plant Biochem. Physiol.and Mol.Biol. "Phosphorylation - Dephosphorylation of Plant Proteins. Abstr. poster 1-63 p. 144

942/A DOBROWOLSKA G, SZCZEGIELNIAK J., MIKOLAJCZYK M, GAGANIDZE D., JURKOWSKI I, MUSZYNSKA G. Salt stress responsive protein kinases from maize seedlings. Ibid.: poster 1-62 p. 143

943/A AWOTUNDE O.S., ZOLNIEROWICZ S., MUSZYNSKA G Identification of protein phosphatase type 2A (PP2A) 65 kDa regulatory subunit in maize seedlings. Workshop:"the Structural Basis for the Phosphate Signaling". Tatra Mountains 20-26 May, 1996 Zakopane, Poland Abstr. 28

944/A MIKOLAJCZYK M., SZCZEGIELNIAK J., GAGANIDZE D., JURKOWSKI I, DOBROWOLSKA G., MUSZYNSKA G. Protein kinases involved in stress signaling in plants. Ibid.: 30

945/A GORA A., PAWLOWICZ J., KIERZEK A., CANDRESSE T., ZAGORSKI W. Pedigree analysis of potato spindle tuber viroid genome. Xth International Congress of Virology. Jerusalem, Israel, August 11-16, 1996 Abstr. PW09 - 5

946/A CHACHULSKA A. M., ROBAGLIA C, CHRZANOWSKA M., ZAGORSKI W. Tobacco veinal necrosis determinants and group specific epitopes in PVY genome. Ibid.: PW12-1

118 947/A SWIEZEWSKA E. Prenylation and acylation of proteins (in Polish). XXXIInd Meeting of Polish Biochemical Society, 17-20 September, 1996 Krakow Abstract VI-4 p. 161, Plenary lecture

948/A HULAMCKA D., JUSZCZUK M. Expression of genes located on the subgenomic RNA of potato leafroll virus. Xth International Congress of Virology 11-16 August 1996, Jerusalem. Abstr.: PW50-5, p.235

949/A SADOWY E., GRONENBORN B., HULAN1CKA D. Infectious transcripts of potato leafroll virus. Ibid.:PWll-18, p.133

950/A SIWECKA M. A. Rye germ ribosomal nuclease degrading double-stranded RNA: purification and specificity. "Ribonucleases: Chemistry, Biology, Biotechnology." 4th Intern. Meeting, Groningen, July 14-18, 1996. Abstr.S2 P4

951/A MICHALIK J., SZOLAJSKA E. Baculoviruses-alternative to chemical pesticides (in Polish). Summer School on ,,Chemistry and Biological Activity of Pesticides" Wroclaw University, Ladek Zdrqj 24-28 June 1996 Abstr.: p. 3

952/A PRZEWLOCKI G., EDELMAN A., PRZYKORSKA A. New sequence specific human Rnase. "Ribonucleases: Chemistry, Biology, Biotechnology." 4th Intern. Meeting, Groningen July 14-18, 1996. Abstr.: S3 Pll

953/A HULANICKA D., PALUCHA A., SADOWY E., ZAGORSKI W. Potato leafroll virus: the engineering resistance to this virus and mechanisms of its genome expression. The second Japonase-Polish Seminar on Biotechnology. Organized by JSPS and PAS "Plant Biotechnology" 23-25 October 1996. Osaka, Japan . Abstr. p. 10

954/A FIKUS M., PAWLOWSKI P. Fatigue of the cellular membrane of Neurospora crassa ( Slime) cells subjected to mechanical stress. 6th Conf. Cell Biology. 12-14 September, 1996 . Lublin,Poland. Folia Histochem. Cytobiol. (1996) 34 suppl. 2 p. 9

119 955/A PALUCHA A, CHRZANOWSKA M, ZAGORSKI W., HULANICKA D. Construction of transgenic lines of potato resistant to potato leafroll virus (in Polish). Seminar: ,,Molecural Basis of Plant-Pathogen Interaction" 14.11.1996, Poznan. Institute of Plant Genetics, PAS

956/A JUSZCZUK M, HULANICKA D. The expression of genes located on the subgenomic RNA of potato leafroll virus (in Polish). Ibid.

957/A HULANICKA D., PALUCHA A, SYLLER J, CHRZANOWSKA M. A transgenic potato carrying cDNA of potato leafroll virus coat protein (in Polish). XXIXth Scientific Session, 5-6 March 1996. Ojrzanow "Genetic-Breeding Bases of Biological Progress in Potato Production", The Potato Institute, Bonin. Abstr. p. 16-17

958/A CHRZANOWSKA M., CHACHULSKA A.M. Advances in the knowledge on variability of PVY strains (in Polish). Ibid.: p.25

959/A CHRZANOWSKA M., CHACHULSKA A. Composition of potato virus Y population described recently in Poland. Characteristics of isolates (in Polish). Ibid.: p.49

960/A JANION C. The MFD effect and DNA repair in E.coli K-12 strains. 5 International Conference on Mechanism of Antimutagenesis and Anticarcinogenesis (5 ICMAA) December 2-6, 1996, Okayama, Japan Abstr. 2-6 p.33

96I/A MOSPAN O., ZASTAWNY T.H., TUDEK B. DNA repair enzyme activity and oxidative DNA damage in the tumor and non-tumorous tissues from lung cancer patients (in Polish). IInd Conf. "Recent Trends in Biochemistry and Biotechnology" Lodz, 12-13 December 1996 Abstr. 111.77

962/A GRA_ZIEWICZ M. A., LAVAL J., TUDEK B. Imidazole ring-opened purines as potentially premutagenic lesions. "DNA Repair Network Workshop" Cancer Res. Inst. SAS, Smolenice Castle, Slovakia 29 April - 1 May, 1996 Abstr. L4, P27

120 963/A GRAZIEWICZ M. A., ZASTAWNY T. H., TUDEK B. Thymine glycols are more potent inhibitors of DNA replication than imidazole ring opened purines (in Polish). IInd Conf. "Recent Trends in Biochemistry and Biotechnology" Lodz, 12-13 December 1996 Abstr. 111.88

964/A CHACINSKA A., BOGUTA M. Interaction of yeast mitochondrial ribosomal protein Nam 9 with chaperone Hsp 104. 24th Intern. Conf. FEBS, Barcelona July 10-12 1996 Abstr. 27

965/A MIECZKOWSKI P., FIKUS M., CIESLA Z. Characterization of a novel gene of Saccharomyces cerevisiae, DIN7, the expression of which is induced by DNA damage. "DNA Repair Network Workshop" Cancer Res. Inst. SAS, Smolenice Castle, Slovakia 29 April - 1 May, 1996, Abstr. P 3 p.37

966/A ADAMCZYK M., SLEDZIEWSKA-GOJSKA E. Pol2-RAD53-DUN1 regulatory pathway is not involved in damage-inducible expression of the Saccharomyces cerevisiae MAG gene. 26 EEMS Annual Meeting. Workshop on chromosome instability and cell cycle control. September 3-7, 1996 Rome, Abstr. p.80

967/A KLUDKIEWICZ B., GODLEWSKI J., GRZELAK K., CYMBOROWSKI B. Effect of ecolysteroid on the synthesis of some proteins in diapausing Galleria larvae. XIIth Ecdysone Workshop Dept. of Agrobiology (CID, CSIC) Jordi Girona 18, Barcelona, July 22-26, 1996, Abstr. 69

968/A EJCHART A., S1EDLECKA M., STICHT H., BIERZYNSK1 A, NMR Studies of secondaiy structure in calcium binding peptide AcDKDGDGYISAA-EAAAQNH2. XIIIrd European Experim. NMR Conference, Paris, 19-24 May 1996 Abstr. p.252

969/A EJCHART A., SIEDLECKA M., STICHT H., BIERZYNSKI A.. Secondary structure of calcium binding peptide AcDKDGDGYISAAEAAAQNH2 International Conference on Magnetic Resonance in Biological Systems. Keystone, USA, August 18-23, 1996

121 970/A CEGLOWSKI P, PANKIEWICZ R., AYORA S, ALONSO J. C. Regulation of gene expression in the regions involved in stable maintenance of pSM 19035 ESF Network Workshop "Molecular Biology and Ecology of Plasmid-mediated Gene Transfer. II Workshop. June 28- July 1, 1996 Potsdam, Germany, Abstr. p.34

97 I/A TOCZYLOWSKA B., ZHUKOV I., DE.BICKI G. High resolution H NMR spectroskopy of human cerebrospinal-fluid; preliminary study (in Polish). XXIX1'1 Seminar on: ,,Nuclear Magnetic Resonance and its Application" Krakow 2-3, December 1996, Poster No. 72

972/A BRETNER M., FELCZAK K., DRABIKOWSKA A., POZNANSKI J., KRAWIEC K, RODE W, PIASEK A., SHUGAR D., KULIKOWSKI T. Thiated pyrimidine deoxynucleoside analogues, potential chemotherapeutic agents, and substrates/inhibitors in various enzyme systems. XII International Roundtable "Nucleosides, Nucleotides and Their Biological Applications", "Making Drugs Out of Nucleosides and Oligonucleotides" September 15-19, 1996 Hyatt Regency, La Jolla, California USA Abstr.p.99

973/A KLECZKOWSKIK. Potato transgenic plants (in Polish). IInd Conference dedicated to prof. A. Dmochowski. Lodz, 12-14 December 1996 Abstr. p. 144-146

974/A KRZYMOWSKA M., TALARCZYK A, HENNIG J. Role of salicylic acid in the response of potato plants to PVY infection (in Polish). Seminar:,,Molecural Basis of Plant-Pathogen Interaction" 14. 11.1996, Poznan. Institute of Plant Genetics, PAS

975/A GORA-SOCHACKA A., CANDRESSE Th, ZAGORSKI W. Single point mutations in the RNA of potato spindle tuber viroid (PSTVd) leading to phenotypical conversions (in Polish). Ibid.

976/A PAWLOWICZ J, GORA -SOCHACKA A., CANDRESSE Th, ZAGORSKI W. Influence of structure of the P domain of potato spindle tuber viroid (PSTVd) on the severity of infection symptoms (in Polish). Ibid.

122 9111A LIPSKA A., CHACHULSKA A.M., ROBAGLIA Ch., ZAGORSKI W. Introduction of cDNA coding for the PVY replicase into the plant genome is one of strategies of getting plants resistant to virus infections (in Polish). Ibid.

978/A HUG G. L., MARCINIAK B, BOBROWSKI K. Acid-base equilibria involved in secondary reactions following the carboxybenzophenone sensitized photooxidation of methionyl-glycine in aqueous solution. Spectral and time resolution of the decaying (S .\N)+ radical cation. Proc. of the Twentieth Doe Solar Photochemistry. Res. Conference French Lick Springs Conf. Center, French Lick, Indiana, June 8-12, 1996. Poster 21, p.119

979/A BANECKI B., MARSZALEK J., WAWRZYNOW A., LIBEREK K., BLASZCZAK A., BARSKI B., JAKOBKIEWICZ J., OGRODNIK D., ZYLICZ M. The role of various molecular chaperones in DNA replication and proteolysis. EMBO Meeting: Molecular biology of DNA replication. Weggis, Suisse, September 8-13 1996, Abstr. p.23

980/A. BARDOWSKI J., LEWICKA K. Modern methods of identification and differentiation of dairy starters (in Polish). VIth Scientific Session on ,,Progress in Technology and Organization in Dairy Industry" Olsztyn 20-21 February 1997 Abstr. p.126-129

98I/A. LIBUDZISZ Z., WALCZAK P., BARDOWSKI J. Lactic acid bacteria - classification, metabolism, genetics, applications (in Polish). Biotechnologia (1997) I (36), 167-171

982/A. BIERZYNSKI A. Structural investigations of EF-hand proteins and their fragments (in Polish). IVth Symp. on Bioorganic and Biomedical Chemistry. Karpacz, 5-8 June 1997 Abstr. p. 10

983/A. TREMBACZ H., JEZEWSKA M. M. Adenine nucleoside phosphorylases in F. hepatica, the mammalian parasite. 9th International/ 6th European Joint Symposium on Purine and Pyrimidine Metabolism in Man. Clinic Biochem.(l997) 30, 20 Abstr.79

984/A. CZERNIEC E., BARDOWSKI J. Identification and characterisation of the amylolytic strain of I. lactis. 8 ameColloque du Club des Bacteries Lactiques. Dijon, 28-30 May 1997 Abstr. Aff.P3

123 985/A. SZYMANSKA M., SWIEŻEWSKA E. Prenylation of Rab 3a protein in vitro. W. Mejbaum-Katzenellenbogen' s Seminars on Molecular BiologyANuclear Organization and Intracellular Transport (ARF and protein 14-3-3), June 23-25, 1997 Wrocław, Szklarska Poręba Abstr. Cell. Mol. Biol. Lett, p.246

986/A. SWIEŻEWSKA E. Prenylation of proteins. Ibid.: Abstr. Plenary lecture

987/A DZIK J. M., GOŁOS B., ZIELIŃSKI Z., RODE W., BRETNER M., FELCZAK K., KULIKOWSKI T. N4-hydroxy-5-halogeno-2'-deoxycytidines and their 5'-monophosphates as inhibitor of thymidylate synthase and in vitro antileukemic agents. 9th Intern./6th European Joint Symp.on Purine and Pyrimidine Metabolism in Man. Clinical Biochem. (1997) 30 24 Abstr. 96

988/A KULIKOWSKA E ., BZOWSKA A., HOLY A., SHUGAR D. Antiviral phosphonate analogs as inhibitors of purine nucleoside phosphorylase. Ibid.: Abstr. 95 p.24

989/A TOBIASZ A, ŻOŁĄDEK T. mdpl mutations located in the 3-end of RSP5 gene encoding ubiquitin-protein ligase are suppressed by mutations in PMA1, a plasma membrane H+ - ATPase gene. 18th Int.Conf. on Yeast Genetics and Molecular Biology. Stellenbosch, South Africa March 31-April 5, 1997 p.175

990/A. KAMIŃSKA J., GAJEWSKA B., ŻOŁĄDEK.T. RSP5 ubiquitin-protein ligase and PAN IP involved in actin cytoskeleton organization interact in two-hybrid system. 9"'Tenovus-Scotland Symposium, Glasgow, April 9-11, 1997 poster 1

99l/A. KONOPIŃSKA A., SZCZEŚNIAK B., BOGUTA M. Genetic interaction between GDS1 and HSP104 of Saccharomyces cerevisiae. Ibid.: poster 2

124 992/A. FELCZAK K., BRETNER M., BARTUZI K., BALINSKA M., KULIKOWSKI T. Synthesis and biological properties of 2'-C-hydroxymethylribose (hamamelose) nucleosides. 9th European Carbohydrate Symposium - Utrecht 1997

July 6-11, 1997 Abstrcts eurocarb9 A169 p.249

993/A. KURLANDZKA A, RYTKA J., RÓŻALSKA B, WYSOCKA M. An essential Saccharomyces cerevisiae protein indirectly affecting cell- substratum interactions. FEBS 97 Special Meeting „Cell Signalling Mechanisms", Amsterdam, July 1, 1997. Abstr. P 6-015

994/A. ZARĘBSKA Z., WASZKOWSKA E., POZNAŃSKI J. Photoadducts of lecithins: their possible role in the phototherapy PUVA (psoralen + UVA). 5th International Student's Scientific Conference, Gdańsk, May 1997 Abstr. 46

995/A. TREMBACZ H., JEŻEWSKA M.M. Deamination of adeninę nucleosides in snails belonging to Helicidae family (in Polih). XXXIIIrd Meeting of Polish Biochemical Society. Katowice, 9-12 September 1997, Abstr. VIII-P16

996/A. TOMASZEWSKI M., WIEJAK J., SIWECKA M. A. Dimeric structure and activity of rye germ ribosomal nuclease degrading double-stranded RNA (in Polish). Ibid.: VIII-P32

997/A. ZARĘBSKA Z., WASZKOWSKA E., CAFFIERI S., DALL'ACQUA F. Psoralen photolesions in the phospholipids. First Internet Conference on Photochemistry and Photobiology November 17 - December 12, 1997 http://www.netsci-journal.com/97v3/

998/A. BARDOWSKI J., TAILLIEZ P., CHOPIN A. Genetic diversity of the lactococcal strains isolated from their natural niche in Poland. An International Symposium on Lactic Acid Bacteria organized by Adria Normandie, the TNO and the Caen University, 10-12 September 1997 Abstr. p. 192

999/A. CHACHULSKA A. M., LIPSKA-DWUŻNIK A., ROBAGLIA, Ch., CHRZANOWSKA M., FLIS B., ZAGÓRSKI W. Potato virus Y genes and proteins, perspectives for engineered plant resistance. Plenary Lecture Biol. Bull. Poznan (1997) 34 (Suppl.): 95

125 1000/A. SKORUPIŃSKA K., SZYMAŃSKA M., HERTEL J., CHOJNACKI T. The occurence of the longest chain polyprenols in leaves of plants. 38th International Conf.on the Biochem.of Lipids, September 16-19, 1997, Assisi, Italy Chem.Phys.Lipids (1997) 88. 141 Abstr.T4/18

100I/A. SZCZERBAKOWA A., MACIEJEWSKA U., BORKOWSKA M., WIELGAT B. Somatic hybridization between Solarium nignim and Solarium tuberosum. Plenary Lecture Biol. Bull. Poznan (1997) 34 (Suppl.): 107-108

1002/A. TALARCZYK A., KRZYMOWSKA M., HENNIG J. Expression of yeast catalase gene in tobacco. 3rd Intern. Conf. „Oxygen, free radicals and environmental stress in plants", Pisa, 15-18 September 1997 Scuola S. Anna, Via Carducci 40 Abstr. p.55

1003/A. KRZYMOWSKA ML, TALARCZYK A., HENNIG J. Is tobacco response to TMV infection modulated by catalase activity? Plenary Lecture Biol. Bull. Poznan (1997) 34 (Suppl.): 102

1004/A. TUDEKB. Formation of imidazole ring-opened purines in DNA and their biological consequences. Conf. on:"Mechanisms of DNA repair and mutagenesis", Warsaw, October 8-11, 1997, Abstr. p.5

1005/A. FIJAŁKOWSKA I. J. Unequal fidelity of leading and lagging strand replication on Escherichia coli chromosome. Ibid.:p.l5

1006/A. BĘBENEK A. A new gene in E. coli K12, isfA, may be involved in down regulation of the SOS system. Ibid.: p.22

1007/A. PIETRZYKOWSKA I., CZAJKOWSKA A. Studies on UV-induced mutagenesis not related to DNA replication. Ibid.: p.23

1008/A. JANION C, WÓJCIK A. Effect of transposon Tn 10 on the survival and mutation frequency of halogen light-irradiated E. coli K-12 strain AB1157. Ibid.: p.24

126 1009/A. KUSMIEREK J. T., BUKOWSKA A.M. Template-directed base pairing of 2-chloro-2'-deoxyadenosine catalyzed by AMV reverse transcriptase. Ibid.: p.27

1010/A. ADAMCZYK M., NOWICKA A., SLEDZIEWSKA -GOJSKA E. Transcriptional induction of the Saccharomyces cerevisiae MAGI gene, in response to DNA-damaging agents, requires neither the MEC1, POL2, RAD53 nor DUN1 regulatory gene products. Ibid.: Poster nr 10 p.47

101 I/A. BORYS E., KUSMIEREK J.T. Endogenous and exogenous DNA lesions recognized by N-alkylpurine-DNA glycosylases. Ibid.: Poster nr 20 p.51

1012/A. FIKUS M. U., RYTKA J., CIESLA Z. Inactivation of a protein kinase encoded by DUN1, that controls the DNA damage response in Saccharomyces cerevisiae, results in a high frequency of rho'mutants. Ibid.: Poster nr 12 p.55

1013/A. GAWEL D., JONCZYK P., SCHAAPER R. M., FIJA1KOWSKA I. J. The assymetry of mutagenesis during replication of leading and lagging DNA strands in E. coli chromosome. Ibid.: Poster nr 1 p.57

1014/A. GRAZIEWICZ M. A., ZASTAWNY T. H., OLINSKI R., SIEDLECKI J., TUDEK B. Inhibition of DNA sythesis on templates oxydized by hydroxyl radical generating system. Ibid.: Poster nr 21 p.58

1015/A. GRZESIUK E., GOZDEK A., NOWOSIELSKA A. Is the DNA polymerase III involved in the preferential transcription Coupled DNA repair ? ' y Ibid.: Poster nr 4 p.60

1016/A. HALAS A., BARANOWSKA H., POLICINSKA Z., JACHYMCZYK W Influence of nuclear DNA polymerases and MSH3 gene on adaptive mutations in yeast Saccharomyces cerevisiae. Ibid.: Poster nr 13 p.61

127 1017/A. JONCZYK P., NOWICKA A., FIJALKOWSKA I., SCHAAPER R. M., CIESLA Z. In vivo protein interactions within the E. coli DNA polymerase III core. Ibid.: Poster nr 3 p.63

1018/A. KRASZEWSKA E., DOBRZANSKA M., TUDEK B. Repair of alkylation DTSIA damages in BY-2 tobacco cells. Ibid.: Poster nr 14 p.67

1019/A. MIECZKOWSKI P., FIKUS M.U., KOPROWSKI P., CIESLA Z. DIN7, a DNA damage-inducible gene of Saccharomyces cerevisiae, is a close homolog of the £TO/gene involved in DNA mismatch repair. Ibid.: Poster nr 11 p.70

1020/A. SPE1NA E., CIESLA J., JANION C. Protecting effect of UmuD' on plasmid DNA. Ibid.: Poster nr7 p.79

1021/A. MALISZEWSKA TKACZYK M., JONCZYK P., PODSIADLA M., SCHAAPER R. M., FIJALKOWSKA I. J. The difference in the fidelity of DNA synthesis between leading and lagging strands replication in E. coli strains. Ibid.: Poster nr 2 p.81

1022/A. WOJCIK A., JANION C. Different responses of E. coli AB1157 strains bearing TN70, TN10KAN and TN9 transposons after halogen light and UV light irradiation. Ibid.: Poster nr 6 p.83

1023/A CHACINSKA A., BOGUTA M. Effect of the cytosolic chaperone HsplO4p on the mitochondrial protein Nam 9p in Saccharomyces cerevisiae. 18 th Int. Conf. on Yeast Geneticis and Molecular Biology. Stellenbosch, South Africa, March 31-April 5, 1997 Abstr. P76 SI29

1024/A TOBIASZ A., ZOLADEK T. mdp I mutations located in the 3 -end of RSP5 gene encoding ubiquitin- protein ligase are suppressed by mutations in PMA1, a plasma membrane H+- ATPase gene. Ibid.: P 114 S175

1025/A ZAGULSKI M, RYTKA J., HERBERT C.J. A functional analysis of YBR142W, an essential gene which is allelic to MAK5-1 and encodes a dead-box helicase. Ibid.: P 182 S255

128 1026/A RYTKA J., KUCHARCZYK R, ZAGULSKI M., HERBERT C.J. The gene YJRO25C encodes the yeast 3-hydroxyanthranilic dioxygenase. Ibid.: P 192 S265

1027/A GROMADKA R., KANIAK A., SLONIMSKI P.P., RYTKA J. The expression of the essential gene KRR1 (YCL059c) is dependent on gene dosage and growth conditions. Ibid.: P 193 S266

1028/A CHACHULSKA A. M., ROBAGLIA Chr., CHRZANOWSKA M., ZAGORSKI W.: Determination des traits necrotiques et sequences des regions 3 terminaies du virus PVY. 16cmes Rencontres de Virologie Vegetale 3-13 March 1997 Del. Region. Alsace du CNRS. Abstr. p.73

1029/A PRZYKORSKA A, YIN Y, KULIGOWSKA E, CARTER C. W. The evidence of conformational changes in tRNATip upon binding with aminoacyl-tRNA synthetase. 17th Intern. tRNA Workshop Kazusa Akademia Park, Chiba, Japan Abstr. 4-23

1030/A TOBIASZ A., HOPPER A. K., ZOLAJDEK T, Mutations located in the 3 -end of RSP5 gene encoding ubiquitin- protein ligase and its suppressors. Mid-Atlantic Yeast Meeting, 19,20,21 June Pittsburhg, PA Abstr. p.28

103 1/A KULCITSKI V., JATMKOWSKI W., CHOJNACKI T., VLAD P. F., UNGURN. D. Superacidic cyclization of polyprenoic acids methyl esters. XVII Conf. on Isoprenoids, 21-26 September, Krakow, Poland 1997

1032/A LIPSKA A., CHACHULSKA A. M, ZAGORSKI W, Molecular analysis of the PVY polymerase gene and its use for plant transformation. 5lh ISSC 22-24 May Gdansk, Poland 1997, Abstr. p.33

1033/A CHECHLACZ M., MICHALIK J., CYMBOROWSKI B. Brain response to suboptimal temperature stress in Galleria mellonella larvae. II-nd Intern. Conf. on Insects, Chemical Physiol. and Environmental Aspect. 14-19 September La_dek Zdroj Abs. P 15

1034/A CHECHLACZ M., MICHALIK J., CYMBOROWSKI B. Effect of suboptimal temperature stress on the profile of brain proteins in Galleria mellonella larvae. Ibid.: P 16

129 1035/A GRZESIUK E., JANION C. Does the mismatch repair system influence the transcription coupled preferential DNA repair. 7th Internat. Conf. on Environmental Mutagens Toulouse, France, 7-12 Sept. 1997 Mutation Res. (1997) J79 suppl. S37 Abstr. PV.14

1036/A JANION C, FABISIEWICZ A. On UV-induced MFD phenomenon in AB1157 E.coli K-12 strains. Ibid.: PV.16 p.S37

1037/A WOJCIK A., JANION C. Mutation frequency decline and DNA repair in halogen light irradiated Escherichia coli K-12 strains. Ibid.: PV.17 p.S38

1038/A. GRAZIEWICZ M. A., ZASTAWNY T. H., TUDEK B. Imidazole ring-opened purines - They are weaker inhibitors of DNA replication than oxidative derivatives of pyrimidines. Ibid.: Poster P XVII B.I p. S172

1039/A TUDEK B. Mutagenic properties of 7-methyladenine. Ibid.: Poster P XVII B.7 p.S174

1040/A. GRAZIEWICZ M. A., ZASTAWNY T. H., TUDEK B. DNA replication on templates modified with hydroxyl radicals generating system-hypoxantine-xanthine oxidase. Vth Intern. Students Scientific Conference 1997 Gdansk, Poland Abstr.p.27

1041/A. SPEINA E., CIESLA J, JANION C. Protecting effect of UmuD' on plasmid DNA integrity. Ibid.: p.42

1042/A. WOJCIK A, JANION C. Effects of halogen light irradiation in Escherichia coli K-12 AB1157 strains. Ibid.: p.47

1043/A. ZIELENKIEWICZ U.( CEGLOWSKI P. Genes epsilon and zeta of pSM19035 act as a plasmid addiction system. 3rd Workshop ,,Molecular Biology and Ecology of Plasmid -Mediated Gene Transfer" 12-16 September, Cuenca (Spain) Abstr. p.36

1044/A. PRYMAKOWSKA-BOSAK M., PRZEWLOKA M., IWKIEWICZ J., EGIERSZDORFF S., GIGOT C, CHAUBET N., KURACE M., SPIKER U., JERZMANOWSKI A. Molecular analysis of transgenic tobacco bearing histone HI gene from

130 Arabidopsis thaliana. 8tK I.M.P. Spring Conference ,,Genes" 22nd to 24th May 1997, Austria Center Vienna Abstr. Poster 72 p.121

1045/A. CALIKOWSKI T., KOZBIAL P., JERZMANOWSKI A. The study of phosphorylation of histon HI with transgenic Nicotiana tobacum plants and the N. tobacum cell line BY-2. Ibid.: Poster 9 p.58

1046/A. BRZESKI J., PODSTOLSKI W, OLCZAK K., JERZMANOWSKI A. Bushy growth : the Arabidopsis thaliana homolog of a component of the yeast SWI/SNF complex takes part in plant morphogenesis. Ibid.: Poster 7 p.56

1047/A. TOMASZEWSKI R., JERZMANOWSKI A. The AT-rich flanks of the oocyte-type 5S RNA gene of Xenopus laevis act as a strong local signal for histon HI-mediated chromatin reorganization in vitro. Ibid.: Poster 93 p. 142

1048/A. PRYMAKOWSKA-BOSAK M., PRZEWLOKA M, OLEJNIK A., KILJANCZYK B., SLUSARCZYK J., KURAS M., JERZMANOWSKI A. Analysis of transgenic tobacco with drastically reduced level of histone HI. EMBO Workshop :"Chromatin and Epigenetic Regulation „ Heidelberg, October 4-7, 1997,Abstr. p. 148

1049/A. MITTELSTEN SCHEID O., PRYMAKOWSKA-BOSAK M., JERZMANOWSKI A., PASZKOWSKI J. Generation of epialleles with paramutation-like interaction by polyploidization of Arabidopsis thaliana. Ibid.: p.135

1050/A. CALIKOWSKI T., KOZBIAL P., JERZMANOWSKI A. Studies of histone HI phosphorylation throughout the cell cycle of tobacco BY-2 cells and during development of the tobacco plants. Ibid.: p.104

105 I/A. GODLEWSKI J, KLUDKIEWICZ B., GRZELAK K., CYMBOROWSKI B. Expression of genes encoding larval hemolymph protein in diapausing Galleria larvae. IInd Intern.Conference on Insects Chemical,Physiological and Environmental Aspects, 14-19 September 1997, La_dek Zdrqj, Poland Abstr.P 14

131 1052/A PIETRZYKOWSKA I., CZAJKOWSKA A. Effect of mismatch repair on UV- and MMS-mutagenesis of 1 susO8 phage induced under conditions permissive and nonpermissive for phage DNA replication. 7th International Conference on Environmental Mutagens.Toulouse, 7-12 September 1997. Mutation Res. (1997) 379 Suppl. SI8 PHI A.5

1053/A. BEJBENEK A., PIETRZYKOWSKA I. isfA mutation inhibits UV-induced GC->AT and AT->GC transitions in recA' strains and GC->TA and AT->TA transversions in recA730 strains. Ibid.: S52 P VII. 22

1054/A. MIKOLAJCZYK M., AWATUNDE O.S ., DOBROWOLSKA G., MUSZYNSKA G. MAP kinases involved in salinity stress response in plants FEBS Special Meeting 1997 ,,Cell Signalling Mechanisms" June 29-July 3, Amsterdam, The Netherlands Abstr. PI-116

1055/A MIKOLAJCZYK M, AWATUNDE O S., MUSZYNSKA G, DOBROWOLSKA G. Protein kinases activated by salinity stress in plants. An ASBMB Satellite of the International Congress of Biochemistry ,,Cellular Regulation by Protein Phosphorylation: 40 Years of Progress" August 20-23, 1997 Seattle, Washington Poster session nr 12: P12 -9

1056/A. TROJANEK J., DOBROWOLSKA G., MUSZYNSKA G. Evidences for existence of protein tyrosine kinases in maize seedlings 8th International Center for BioTechnology Symposium on Cytoplasmic Protein -Tyrosine Kinases. May 23-25 1997, Karolinska Institutet NOVUM Research Park, Huddinge, Sweden Abstr. Poster P 16 p.64

1057/A. DOBROWOLSKA G., MIKOLAJCZYK M., SZCZEGIELNIAK J., AWOTUNDE O.S., TROJANEK J., MUSZYNSKA G. Reversible phosphorylation of proteins in plant salinity stress response Biol. Bull. Poznari (1997) 34 (Suppl.), 28

1058/A. CHACHULSKA A. M. Molecular analysis of potato virus Y genome in Poland (in Polish). Scientific Conference: ,,Viral Potato Diseases - New Trends in Research" 4-5 April 1997, Ojrzanow-Mlochow, Bonin 1997 Abstr.p. 8

132 1059/A. CHACHULSKA A. M., CHRZANOWSKA M., FLIS B., ROBAGLIA Ch., ZAGÓRSKI W. Antisense and sense RNA-mediated resistance to potato virus Y in potato and tobacco. 5th International Congress of „Plant Molecular Biology" 21-27 September 1997 (SICEC) Singapore Plant Mol. Biol. Reporter (suppl.) (1997) 15, Abstr.1392

1060/A KRUSZEWSKA J. S., BUTTERWECK A. H., MIGDALSKI A., KUBICEK Ch., PALAMARCZYK G. Expression of the Saccharomyces cerevisiae DPMI (mannosylphosphodolichol-synthase-encoding) gene in Trichoderma reesei results in the increased level of protein secretion. TRICEL 97 „Carbohydrases from Trichoderma Reesei and other Microorganisms .Structures/Biochemistry Genetics/Applications Ghent-Belgium, August 28-30, 1997 Abstr. P 41

106I/A. KRUSZEWSKA J. S., SALOHEIMO M., PENTTILA M., PALAMARCZYK G. Isolation of the Trichoderma reesei c-DNA that encodes GTP:a-D-mannose- 1 -phosphate guanyl transferase and functions in Saccharomyces cerevisiae. Ibid.: P 45

1062/A. SZKOPIŃSKA A., GRABIŃSKA K., DELOURME D, KARST F., PALAMARCZYK G. Farnesyl diphosphate synthase activity and polyprenol and dolichol formation in the yeast Saccharomyces cerevisiae. 3ld TERPNET Meeting of the European Network on Plant Isoprenoid „Isoprenoid Biosynthesis Regulation and Biotechnological Applications" Univ. de Poitiers, France, May 29-30, 1997 Abstr.

1063/A. PASZEWSKI A., LEW ANDO WSKA I., BALIŃSKA M. Genetic regulation of folate enzymes synthesis in Aspergillns nidulans. 11th International Symposium on Chemistry and Biology of Pteridines and Folates. Pteridines ( 1997) 8, 123

1064/A. NATORFF R., PIOTROWSKA M., PASZEWSKI A. The Aspergillns nidulans sconB sulphur regulatory gene encodes a protein with WD40 repeats and F-box. 19th Fungal Genetics Conference.March 18-23, 1997 Asilomar Conf.Center Pacific Grove California Abstr.Poster nr 36 p. 96

1065/A. PIOTROWSKA M., NATORFF R., PASZEWSKI A. SconC, an Aspergillus nidulans sulphur metabolism regulatory gene is homolog of yeast SKPI essential gene. Ibid.: Poster nr 39 p.97

133 1066/A. HULANICKA M. D. Non-convention strategies to protect plants against viral infections. VIIIth Conference of the Polish IAPTC Section and in vitro Culture Section of the Polish Botanical Society: ,,Plant Cultures in vitro in Basis and Applied Research", Krakow, 25-27 August 1997 Abstr.: p. 13

1067/A. PALUCHA A., HULANICKA M. D. 3. Construction and analysis of potato plants carrying different fragments of potato leafroll luteovirus cDNA. Association of Applied Biologists and Societe Francaise de Phytopathologie Symposium LUTEOVIRUSES 1997, Cirencester, UK

1068/A. JUSZCZUK M., HULANICKA M.D. 2. The expression of genes located on the subgenomic RNA of potato leafroll virus. Ibid.:

1069/A. SADOWY E., GRONENBORN B., HULANICKA M. D. 4. Infectious transcripts of potato leafroll virus. Ibid.:

1070/A. MUSZYNSKAG. Book reviews:Protein Analysis and Purification .Benchtop Techniques. Rosenberg J. M. Birkhauser 1996 ActaPhysiol.Plant.(1997) 19, 390-391

107I/A. CHACINSKA A., ROSPERT S., SCHATZ G., BOGUTA M. Effect of the cytosolic chaperone Hsp 104 on the phenotype of a point mutant in the mitochondrial ribosomal gene NAM9. Biology of Molecular Chaperones. Eurovillage, Obernai, France. May 22-27, 1997

1072/A. HALAS A., BARANOWSKA H., POLICINSKA Z., JACHYMCZYK W. Influence of nuclear DNA polymerases and MSH3 gene on adaptive mutations in yeast Saccharomyces cerevisiae. 7th International Conference on Environmental Mutagens. Toulouse 7-12 October 1997 Mutation Res. (1997) 379, suppl. P III A.10 p.S20

134 1073/A GOLOS B., DZIK J. M, JANKOWSKA J., KRASZEWSKI A., STAWINSKI J., RODE W., SHUGAR D. Synthesis and interactions with thymidylate synthase of the 5'-thiophosphates and 5'-H-phosphonates of 2'-deoxyuridine, thymidine and 5-fluoro - -2'-deoxyuridine. 6th International Symp. on Molecular Aspects of Chemotherapy, Gdansk, Poland, 9-12 July 1997. Abstr. nr 49 p.l 18

1074/A GOLOS B, DZIK J. M., RODE W, FELCZAK K., BRETNER M., SHUGAR D., KULIKOWSKI T. Acyclic analogues of 5-fluoro-dUMP and 5-fluoro-2'- deoxyuridine: Synthesis and inhibition of thymidylate synthase and tumour cell growth. Ibid.: 50 p. 119

1075/A BYKOWSKI T., IWANICKA-NOWICKA R., HRYNIEWICZ M., HULANICKA D. Expression of tauABCD operon representing sulphate starvation inducible genes in Escherichia colt involvement of CysB and Cbl regulatory proteins. 8th European Students Conference of the Charite, Humbold University, Berlin October 15-18, 1997 Abstr. B 68 p. 44

1076/A. JARMULA A., ANULEWICZ R, LES A, CYRANSKI M.K., ADAMOWICZ L., BRETNER M., FELCZAK K., KULIKOWSKI T., KRYGOWSKI T.M., RODE W. Interaction of thymidylate synthase with 5-fluoro-substituted dUMP analogues in view of the pyrimidine ring structure . 6lb Intern.Symp. on Molecular Aspects of Chemotherapy. Gdansk, 9-12 July 1997 Abstr. nr 2 p.68

1077/A. HENN1GJ. Perception of ethylene signal in plant cells. Conf. on Molecular Basis of Signal Transduction in Plants. Warszawa November 28, 1997

1078/A. MIKOLAJCZYK M., AWOTUNDE O. S., DOBROWOLSKA G., MUSZYNSKA G. Protein kinases involved in stress response in plants. Ibid.

1079/A. WE.GRZYNA. Activation of transcription by DnaA protein (in Polish). XXVth Seminar, Institute of Molecural Biology, Jagiellonian University: ,,Molecural Mechanisms of Gene Expression Regulation", Zakopane 22-27 March 1997

135 1080/A. OBUCHOWICZ M., SZALEWSKA-PAŁASZ A, WĘGRZYN A., CIESIELSKA A, GILADI H., THOMAS M.S., OPPENHEIM A. B., WĘGRZYN G. Point mutation in a subunit of E. coli RNA polymerase has many effects on bacteriophage X development. NATO Adv. Study Institute Conf. on Molecular Microbiology, Birmingham England, 4-17 April 1997

108 l/A. BYKOWSKI T., 1WANICKA-NOW1CKA R., HRYNIEWICZ M., HULANICKA M. D. Expression of tauABCD operon representing suphate starvation inducible genes in Escherichia coli involvement of Cys B and Cbl regulatory proteins. 5th International Students Scientific Conference, Gdańsk 10-14 May 1997

1082/A. WANKE M., ŚWIEŻEWSKA E., DALLNER G. Intracellular localization and transport of plastoquinone and ubiquinone in spinach cells. 1st Euroconference on Cell Dynamics. Crete, November 1-4, 1997

1083/A. MIECZKOWSKI P. A., FIKUS M. U., CIEŚLA Z. Regulation of expression of DIN 7, DNA damage-inducible gene of Saccharomyces cerevisiae, which is a structural homolog of the RAD2 and RAD27 DNA repair genes. 18th International Conference on Yeast Genetics and Molecular Biology Stellenbosch, South Africa, March 31-April 5, 1997. Abstr. P 23 p. S69

1084/A ADAMCZYK M., ŚLEDZIEWSKA-GÓJSKA E. The MECÍ, POL2, RAD53, DUN! Saccharomyces cerevisiae regulatory genes do not affect DNA damage-inducible expression of the MAG gene encoding 3-methyladenine DNA glycosylase. 7th International Conf.on Environmental Mutagens, Toulouse, France, 7-12 September 1997 „Fundamental and Molecular Mechanism of Mutagenesis" Mutation Res. A Section 379 supp.l Abstr. P VII. 14

1085/A. FIJAŁKOWSKA I. J., JOŃCZYK P., MALISZEWSKA-TKACZYK M., PODSIADŁA M., GAWEŁ D., SCHAAPER R. M. The assymetry of mutagenesis during replication of leading and lagging DNA strand in E. coli chromosome. Ibid.: P III B.3

1086/A JOŃCZYK P., NOWICKA A., FIJAŁKOWSKA I. J. Specific protein-protein interactions between E. coli DNA replication proteins. Ibid.: P III B.4

136 1087/A. DOBRUK A., KLOPOTOWSKI T. Expression of CSPC gene coding for a cold shock family protein in Escherichia coli K12. Vth International Students Scientific Conference 1997, Gdansk, Poland Abstr. Poster p.23

1088/A. JANOWICZ A, LOBOCKA M. P1 plasmid partition : A method for selection of chromosomal mutants defective in partition dysfunction. Ibid.: p.29

1089/A. FIKUSM. Transgenic plants-transgenic food (in Polish). XXXIII'd Meeting of Polish Biochemical Society, Katowice 9-12 September 1997, Abstr. IV-W3

1090/A FIKUS M., PAWLOWSKI P. Cellular membrane subjected to nonreversible shear deformation. 17th Internat.Congress of Biochemistry and Molecular Biology, August 24-29 1997 S. Francisco USA, Abstr. 753

109 I/A. KOWALSKA A., KLOPOTOWSKI T. Regulatory protein interactions with promoter DNA controling WCA operon coding for exopolisaccharide synthesis and export in Esccherichia coli K-12. Vlh Intern. Students Scientific Conference. Gdansk Poland, 1997. Abstr.p.31

1092/A. RODIONOV O., LOBOCKA M., YARMOLINSKI M. Silencing of genes flanking the PI plasmid centromere. University of Wisconsin-Madison Memorial Union August 5-10, 1997 ,,Molecular Genetics of Bacteria and Phages", Abstr. p. 82

1093/A. TOCZYLOWSKA B., ZHUKOV I. Application of high resolution NMR techniques to study human physiological fluids. Symposium on ,,Application of Magnetic Resonance in Chemistry and Related Areas", June 25-27, 1997, Warszawa, Abstr. P-52

1094/A. ZHUKOV I. Determination of spatial structure of the Cucurbita maxima trypsin inhibitor CMT1 (M8L) by NMR methods. Ibid.: P-60

137 1095/A. WĘGRZYN G., SZALEWSKA-PAŁASZ A., WĘGRZYN A., HERMAN- ANTOSIEWICZ A., BŁASZCZAK A., TAYLOR K. Escherichia coli dnaA function, transcriptional activation of X phage and plasmid replication. Jacques Monod Conference on Regulation of DNA Replication in Prokaryotes and Eukaryotes: Molecular Aspects, Aussois, France

1096/A. WĘGRZYN G., WĘGRZYN A., REICHERT M., TAYLOR K. Replication of À. phage is inhibited in the dnaA mutant devoid of cryptic lambdoid prophages (in Polish). XXXIIIrd Meeting of Polish Biochemical Society, Katowice 9-12 September 1997, Abstr. I-P49 p. 29

1097/A. WĘGRZYN A., TAYLOR K., WĘGRZYN G. Differential sensitivity to rifampicin of Escherichia coli RNA polimerase depending on the kind of a subunit (in Polish). Ibid.: I-P46 p.28

1098/A. SZALEWSKA-PAŁASZ A., WĘGRZYN A., LEMIESZEK E., TAYLOR K, WĘGRZYN G.

Activation of bacteriophage X PR promoter by DnaA protein (in Polish). Ibid.: I-P45 p. 28

1099/A. HERMAN A., WĘGRZYN A., WĘGRZYN G., TAYLOR K. Construction and characterization of the X plasmid devoid of the cro function (in Polish). Ibid.: I-P42 p.27

1100/A. GAB IG M., SZALEWSKA-PAŁASZ A., WĘGRZYN A., OBUCHOWSKI M., KUŹNICKA A, WĘGRZYN G. Production of excess of certain proteins as a developmental strategy of bacteriophage X (in Polish). Ibid.: I-P39 p.26

1101/A WĘGRZYN A., WĘGRZYN G., HERMAN A., TAYLOR K. Escherichia coli cell cycle and regulation of A. plasmid replication Workshop on the bacterial cell cycle, Chorin, Germany 13-17 September 1997

1102/A. WĘGRZYNA. Molecular mechanisms of the disassembly of bacteriophage X replication complex (in Polish). Conference of the Nucleic Acids Section, Polish Biochemical Society, Gliwice 21-22 November 1997

138 1103/A. BLASZCZAK A., ZYLICZ M., GEORGOPOULOS C, LIBEREK K. Autoregulation of response of heat shock in Escherichia coli (in Polish). XXXIIIrd Meeting of Polish Biochemical Society, Katowice 9-12 September 1997, Abstr. I-W12 p. 13

1104/A. BLASZCZAK A., ZYLICZ M., GEORGOPOULOS C, LIBEREK K., The importance of C-terminal part of a32 in FtsH dependent degradation. EMBO Workshop on Cellular Functions of AAA Proteins, Tutzing, 17-21 May.

139 PATENTS

P 4 SKRZECZKOWSKI L. J., ZAGORSKI-OSTOJA W., WELNICKI M. The method of synthesis of polynucleotide probe for detection of potato spindle tuber viroid. Assignee: Institute of Biochemistry and Biophysics PAS, Warszawa Patent Description PL 161561 Bl. Appl. No: 280261, Int Cl5: C12N 15/11 GO1N 33/50

P 5 ZEKANOWSKI C, WELNICKI M., SKRZECZKOWSKI L. J., ZAGORSKI - OSTOJA W. The method of detection of pathogens and diagnostic test for pathogen detection. Assignee: Institute of Biochemistry and Biophysics PAS, Warszawa Patent Description PL 170018 Bl. Appl. No: 288397, IntCl6: C12Q 1/68 C12N 15/10

140 DISSERTATIONS Theses for the degree of ,,doctor habilitatus" (Dr. Sc.)

JARUZELSKA JADWIGA Molecular basis of phenylketonuria: gene mutations and maturation defects of the phenylalanine hydroxylase pre-mRNA. Presented on January 23, 1996. Research done in the Department of Human Genetics PAS, Poznari.

CEGLOWSKI PIOTR Consequences of generation of monomeric plasmid forms in Bacillus subtilis cells. Presented on March 12, 1996. Research done in the Department of Microbial Biochemistry, IBB PAS.

HENNIG JACEK Role of salicylic acid in plant response to pathogen attack. Presented on July 2, 1996. Research done in the Department of Plant Biochemistry, IBB PAS.

KULA-SWIEZEWSKA EWA Role of isoprenoid compounds in plant cells. Biosynthesis, biological function and occurrence. Presented on October 8, 1996. Research done in the Department of Lipid Biochemistry, IBB PAS.

GRZESIUK ELZBIETA Role of Escherichia coli, mutS, umuDC and dnaQ genes in mutagenesis and repair of DNA damages induced by alkylating agents. Presented on December 17, 1996. Research done in the Department of Molecular Biology, IBB PAS.

PRZYKORSKA ANNA RNA structure probing with the use of two plant nucleases. Presented on April 15, 1997. Research done in the Department of Protein Biosynthesis, IBB PAS.

ZIELENKIEWICZ PIOTR Structure prediction and theoretical studies of interactions of small globular proteins. Presented on June 3, 1997. Research done in the Department of Biophysics, IBB PAS.

141 SZKOPINSKA ANNA Biosynthesis of polyprenols and their derivatives.The role of farnesyl diphosphate synthase in common regulation of the level of cyclic and linear polyisoprenoids synthesis. Presented on October 7, 1997. Research done in the Department Lipid Biochemistry, IBB PAS.

Doctoral theses (Ph. D.)

POZNANSKI JAROSLAW Conformational analysis of oligoproline-bridged peptides and its application in modeling of electron transfer between terminal aminoacids (Trp, Tyr, Met). Presented on January 23, 1996. Research done in the Department of Biophysics, IBB PAS.

REMPOLA BOZENNA Synthesis of serine proteinases inhibitor CPTIII in Escherichia coli and Sacharomyces cerevisiae. Presented on March 12, 1996. Research done in the Department of Physics, IBB PAS.

TOPCZEWSKA JOLANTA Synthesis of human epidermal growth factor - hEGF in yeast Saccharomyces cerevisiae. Presented on May 14,1996. Research done in Department of Biophysics, IBB PAS.

TOPCZEWSKI JACEK Cloning and characterization of Aspergillus nidulans cys B gene encoding cysteine synthase. Presented on May 14, 1996. Research done in the Department of Genetics, IBB PAS.

AWAN MALIK FATEH MUHAMMAD Response of potato {Solarium tuberosum L.) tissues to Phytophthora infestans or its elicitors. Presented on June 2, 1996. Research done in the Department Plant Biochemistry, IBB PAS.

PALUCHA ANDRZEJ Agroinfection of selected potato cultivars by binary plasmids carrying fragments of cDNA PLRV genome. Presented on October 8, 1996. Research done in the Department of Microbial Biochemistry, IBB PAS.

142 WIDLAK WIESLAWA Structural and functional analysis of the rat gene hsp70 promoter with the use of the transgenic mouse model. Presented on October 8, 1996. Research done in the Department of Molecular Biology, The Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice.

NOWICKA ADRIANNA Protein-protein interactions between Escherichia coli ,,mutasome" proteins. Presented on December 17, 1996. Research done in the Laboratory of Mutagenesis and DNA Repair, IBB PAS.

KERN IZABELLA Secretion of streptokinase from Streptococcus equisimilis H46A in homologous and heterologous bacterial systems. Presented on February 11, 1997. Research done in the Depertment of Microbial Biochemistry, IBB PAS.

BE_BENEK ANNA A new mutation in Escherichia coli K12, isfA, inhibits SOS dependent mutagenesis and other SOS functions. Presented on February 11,1997. Research done in the Department o Molecular Biology, IBB PAS.

BRETNER MARIA Search of new 2-, 4-, and 2,4-disubstituted pyrimidine nucleosides and nucleotides, potential inhibitors of thymidylate synthase and anticancer agents. Presented on February 11, 1997. Research done in the Department of Molecular Biology, IBB PAS.

SIENKO MARZENA Cloning and characterization of Aspergillus nidulans genes coding for homocysteine synthetizing enzymes. Presented on April 15, 1997. Research done in the Department of Genetics, IBB PAS.

PAWLOWSKI KRZYSZTOF Theoretical modeling of structures of calcium-binding proteins and their fragments. Presented on April 15, 1997. Research done in the Department of Biophysics, IBB PAS.

143 LISOWSKA KATARZYNA Cloning and structure analysis of gene hsp70.1 belonging to a family of rat heat shock genes. Presented on June 3, 1997. Research done in the Department of Molecular Biology, The Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice.

TALARCZYK ELZBIETA Long-chain plant polyprenols; the occurrence and putative role in protein prenylation. Presented on April 15, 1997. Research done in the Department of Lipid Biochemistry, IBB PAS

IWANICKA-NOWICKA ROKSANA New gene of Escherichia coli - cbl. Presented on October 7, 1997. Research done in the Department of Microbial Biochemistry, IBB PAS

Theses for the degree of Master of Sciences (M. Sc.)

TOBIASZ ANNA Genetic, molecular and phenotypic analysis oimdpl mutants and their suppressors. Presented on March 29, 1996. Research done in the Department of Genetics, IBB PAS and the Department of Clinical Biochemistry and Chemistry, the Warsaw Medical Academy

SIEDLECKA MONIKA Structural investigations of a peptide analogous to the third calcium binding loop of calmodulin. Presented on May 21, 1996. Research done in the Department of Chemistry of the Warsaw Politechnical University and the Department of Biophysics, IBB PAS

OLEJNIK ANNA Construction and molecular and fenotypical analysis of tobaco plant Nicotiana tabacum with histone H1:DNA relation disturbed. Presented on May 21, 1996. Research done in the Laboratory of Plant Molecular Biology, Department of Physiology of Plant Resistance, the Warsaw University

RACKI WALDEMAR Analysis of sequence of the omega intron in the SuW Saccharomyces cerevisiae. Presented on May 21, 1996. Research done in the Department of Genetics, the Warsaw University.

144 CHECHLACZ MAGDALENA Changes in the profile of brain proteins as a result of low temperature stress in Galleria mellonella. Presented on June 13, 1996. Research done in the Department of Invertebrate Physiology, Institute of Zoology, the Warsaw University and in theDepartment of Protein Synthesis, IBB PAS.

PALCZEWSKA MALGORZATA Studies on functions of the nuclear genes MSS116 i DSS 1 from Saccharomyces cerevisiae. Presented on July 9, 1996. Research done in the Department of Genetics, the Warsaw University.

GODLEWSKI JAKUB Influence of diapause on the synthesis of some Galleria mellonella larvae proteins. Presented on July 16, 1996. Research done in the Department of Invertebrate Physiology, Institute of Zoology, the Warsaw University and in theDepartment of Protein Synthesis, IBB PAS.

KRYSIAK CEZARY Transient and stable transformation of wheat with the use of a gene gun. Presented on September 27, 1996. Research done in the Department of Plant Biochemistry, IBB PAS and Chair of Plant Physiology, the Warsaw Agriculture University.

NOWOSIELSKA ANETTA Development of 5. cerevisiae expression system for the synthetic chimeric gene CPCI in S. cerevisiae. Presented on September 27, 1996. Research done in the Department of Biophysics, IBB PAS and in the Department of Biology and Earth Sciences, The Maria Curie-Skfodowska Memorial University, Lublin.

TKACZYK MARTA The influence of the potato leaf roll virus subgenomic RNA leader sequence on the reporter gene itidA encoding b-glucuronidase (GUS). Presented on October 9, 1996. Research done in the Department of Microbial Biochemistry, IBB PAS and the Department of Biology, the Warsaw University.

MORAWSKI MARCIN An attempt to clone the regulatory gene sitD of arginine pathway of Aspergillus nidulans. Presented on February 6, 1997. Research done in the Department of Genetics, the Warsaw University.

145 MALEWICZ MICHAL Studies on the function of nuclear genes DSSJ and NUC1 involved in mitochodrial biogenesis in the yeast Saccharomyces cerevisiae. Presented February 6, 1997. Research done in the Department of Genetics, the Warsaw University.

LIPSKA ANNA Molecular analysis of the PVY polymerase gene and its use for plant transformation. Presented on May 9, 1997. Research done in Laboratory of Plant Molecular Biology, the Institute of Plant Experimental Biology, the Warsaw University and in the Department of Protein Biosynthesis, IBB PAS.

KOWALCZYK PIOTR The effect of nuclear and mitochondrial mutations in Saccharomyces cerevisiae on the mitochondrial 21s rRNA omega intron stability. Presented on June 19, 1997. Research done in the Department of Genetics, the Warsaw University.

DZIEMBOWSKI ANDRZEJ Cloning and analysis of the ADZ1 nuclear gene from Saccharomyces cerevisiae. Presented on June 20, 1997. Research done in the Department of Genetics, the Warsaw University.

KACZOR CELINA In vivo interaction beetwen protein products of genes e and x from plasmid pSM19035. Presented on June 24, 1997. Research done in the Department of Microbial Biochemistry, IBB PAS and the Agriculture University.

GAWEL DAMIAN Construction of the chromosomal system to study fidelity of leading and lagging strand DNA replication in E. coll Presented on June 26, 1997. Research done in the Laboratory of Mutagenesis and DNA Repair, IBB PAS and in the Warsaw Agriculture University.

KUSZTYKIEWICZ BERNARDA Photoadducts of psoralen to linoleic acid: isolation and characterization. Presented on July 7, 1997. Research done in the Department of Molecular Biology, IBB PAS and University of M. C. Sklodowska, Lublin.

146 JAWORSKA MALGORZATA Photoadducts of 8-methoxypsoralen with oleic acid. Isolation and characterization. Presented on July 7, 1997. Research done in the Department of Molecular Biology, IBB PAS and University of M. C. Sktodowska, Lublin.

BOROWSKA AGNIESZKA A search for function of a new IRR1 gene of Saccharomyces cerevisiae. Presented on July 7, 1997. Research done in the Department of Genetics, IBB PAS and the Warsaw Agriculture University.

DOBRUK ANETA Expression of cspC gene encoding a cold response protein in E. coli K12. Presented on July 17, 1997. Research done in the Department of Microbial Biochemistry, IBB PAS and University of Gdansk.

KOWALSKA AGNIESZKA Regulatory elements of wca encoding colanic acid synthesis in Escherichia coli K12. Presented on July 17, 1997. Research done in the Department of Microbial Biochemistry, IBB PAS and University of Gdansk.

CZERNIEC EWA Characterization of the amylolytic stain IBB500 of Lactococcus lactis and identification of the amylase encoding gene. Presented on October 8, 1997. Research done in the Laboratory of Plant Molecular Biology, Warsaw University and in the Department of Microbial Biochemistry, IBB PAS.

ALEKSANDRZAK TAMARA New genes of lactose metabolism in Lactococcus lactis. Presented on October 28, 1997. Research done in the Laboratory of Plant Molecular Biology, Warsaw University and in the Department of Microbial Biochemistry, IBB PAS.

ADAMCZYK MALGORZATA Dependence of the expression of MAGI on the POL2, MEC1.RAD53, DUN1 gene products of 5. cerevisiae. Presented on November 17, 1997. Research done in the Laboratory of Mutagenesis and DNA Repair, IBB PAS and the Agriculture University.

147 INSTITUTE RESEARCH STAFF

Note: Names of Research Units are abbreviated as follows: Bi - Biophysics, Ge - Genetics, Inf - Bioinformatics, Li - Lipid Biochemistry, Mi - Microbial Biochemistry, Mo - Molecular Biology, Mu - Mutagenesis and DNA Repair, Nmr - Biological NMR, PI - Plant Biochemistry, Pr - Protein Biosynthesis, Pu - Purine Metabolism, UG - University of Gdansk, UW - Warsaw University.

Baranowska-Wyszomirska Hanna, Ph. D., Ge Bardowski Jacek, Ph. D., Mi Barszcz Daniela, Ph. D., Mo Bartnik Ewa, Prof., UW B^benek Anna, Ph. D., Ge Bierzynski Andrzej, Prof., Nmr Bteszczak Adam, M. Sc., UG Bobrowski Kxzysztof, Dr. Sc., Bi Bolewska Krystyna, Ph. D., Bi Borsuk Piotr, Ph. D., UW Bretner Maria, Ph. D., Mo Brzywczy Jerzy, Ph. D., Ge Bucholc Maria Jolanta, Ph. D., PI Buchowicz Jerzy, Prof., PI Burzyriska Beata, Ph. D., Ge

Cegtowski Piotr, Assoc. Prof, Mi Chachulska Anna, Ph. D., Pr Cheistowska Anna, Ph. D., Ge Chojnacki Tadeusz, Prof., Li Ciesla Jarostaw, Ph. D., Mo Ciesla Zygmunt, Prof., Mu

Dadlez MichaJ, Ph. D., Bi Dmochowska Aleksandra, Ph. D., UW Dobrowolska Grazyna, Ph. D., PI Dobrzanska-Wiernikowska Marta, Ph. D., PI Dominski Zbigniew, Ph. D., Ge Drabikowska Alicja, Assoc. Prof, Mo Dzikowska Agnieszka, Ph. D., UW

Ejchart Andrzej, Dr. Sc., Nmr

Fabisiewicz Anna, Ph. D., Mo Felczak Krzysztof, Ph. D., Mo

148 Fijałkowska Iwona, Ph. D., Mu Fikus Magdalena, Prof., Bi

Gabryszuk Joanna, Ph. D., Pr Gaganidze Dali, Ph. D., PI Goch Grażyna, Ph. D., Bi Grzelak Krystyna, Dr. Sa, Pr Grzesiuk Elżbieta, Dr. Sa, Mo

Hennig Jacek, Assoc. Prof., Pl Hryniewicz Monika, Dr. Sa, Mi Hulanicka-Dziechcińska M. Danuta, Prof., Mi

Iwanicka-Nowicka Roksana, Ph. D., Mu

Jachymczyk Witold, Prof., Ge Jagura-Burdzy Grażyna, Dr. Sa, Mi Janion Celina, Prof., Mo Jankowski Wiesław, Ph. D., Li Jerzmanowski Andrzej, Prof., UW/Pr Jeżewska Maria Monika, Prof., Pu Jończyk Piotr, Ph. D., Mu

Kem Izabela, Ph. D., Mi Kleczkowski Kazimierz, Prof., Pl Kłopotowski Tadeusz, Prof., Mi Kłudkiewicz Barbara, Ph. D., Pr Krajewska-Grynkiewicz Krystyna, Ph. D., Mi Kraszewska Elżbieta, Ph. D., Pl Kroczyńska Barbara, Ph. D., Pl Krwawicz Joanna, M. Sa, Mo Kruszewska Joanna, Ph. D., Li Kula-Świeżewska Ewa, Assoc. Prof., Li Kuligowska Elżbieta, Ph. D., Pr Kulikowski Tadeusz, Prof., Mo Kurlandzka Anna, Ph. D., Ge Kuśmierek Jarosław, Prof., Mo Kwiatkowski Bogusław, Ph. D., Pl

Lasocki Krzysztof, M. Sa, Mo I Lassota Zofia, Prof., Pr | Leszczyńska Krystyna, Ph. D., Mi Lewicka Katarzyna, Ph. D., Mi

149 Łobocka Małgorzata, Ph. D., Mi Łoziński Tomasz, Ph. D., Bi

Maciejewska Urszula, Ph. D., Pl Mazuś Barbara, Dr. Sa, PI Miazga Agnieszka, M. Sa, Mo Michalik Joanna, Dr. Sa, Pr Muszyńska Grażyna, Prof., Pl

Natorff Renata, Ph. D., Ge Nowak Leszek, Ph. D., Isotope Facility Nowicka Adrianna, Ph. D., Mu

Palamarczyk Grażyna, Prof., Li Pałucha Anrdrzej, Ph. D., Pr Paszewski Andrzej, Prof., Ge Pawłowski Krzysztof, Ph. D., Nmr Pawłowski Piotr, Ph. D., Bi Piechocka Maria, M. Sa, Pl Pietrzykowska Irena, Prof., Mo Piotrowska Małgorzata, Ph. D., Ge Płochocka Danuta, Ph. D., Bi Poznański Jarosław, Ph. D., Bi Przewłocki Grzegorz, Ph. D., Pr Przykorska Anna, Ph. D., Pr

Rabczenko Andrzej, Prof., Bi Rakowska-Boguta Magdalena, Ph. D., Ge Rempoła Bożenna, Ph. D., Bi Rytka Joanna, Prof., Ge

Shugar David, Prof. Sieńko Marzena, Ph. D., Ge Sierakowska Halina, Prof., Mo Sirko Agnieszka, Ph. D., Mi Siwecka Maria Agnieszka, Ph. D., Pr Skoneczny Marek, Ph. D., Ge Smagowicz Wiesław, Ph. D., Bi Stańczak Janusz, Ph. D., Pr Stępień Piotr, Prof., UW Szafrański-Szeliga Przemysław, Prof., Pr Szarkowski Jan W., Prof., Pr Szczerbakowa Anna, Ph. D., Pl Szkopińska Anna, Ph. D., Li Szołajska Ewa, Ph. D., Pr Szurmak Blanka, Ph. D., Pl

150 Śledziewska-Gójska Ewa, Ph. D., Mu

Toczyłowska Beata, Ph. D., Nmr Tomaszewski-Chyczewski Marek, Ph. D., Pr Topczewski Jacek, Ph. D., Ge Topczewska Jolanta, Ph. D., Bi Tudek Barbara, Ph. D., Mo

Wasilewska-Dąbrowska Lidia Danuta, Prof., Pl Węgleński Piotr, Prof., UW Węgrzyn Alicja, Ph. D., UG Wielgat Bernard, Assoc. Prof., Pl Wierzchowski Kazimierz Lech, Prof., Bi Wolinowska Renata, Ph. D., Mi Wójcik Jacek, Ph. D., Nmr

Zagulski Marek, Ph. D., Pr Zagórski-Ostoja Włodzimierz, Prof., Pr Zarębska Zofia, Assoc. Prof., Mo Zatyka Małgorzata, Ph. D., Mi Zielenkiewicz Piotr, Assoc. Prof., Inf Żołądek Teresa, Ph. D., Ge Żuk Jerzy, Prof., Ge

School of Molecular Biology:

Adamczyk Małgorzata, M. Se, Mu Aleksandrzak Tamara, M. Se, Mi Białkowska Agnieszka, M. Se, Ge Borys Ewa, M. Se, Mo Brodzik Robert, M. Se, Pl Brzeski Jan, M. Se, Pr Bykowski Tomasz, M. Se, Mi Calikowski Tomasz, M. Se, Pr Chacińska Agnieszka, M. Se, Ge Ciesielski Arkadiusz, M. Se, Ge Czajkowska Anna, M. Se, Mo Czerska Kamila, M. Se, Ge Dobruk Aneta, M. Se, Mi Dyczkowski Jerzy, M. Se, Bi Felczak Magdalena, M. Se, Mo Fikus Marta, M. Se, Mu

Gajewska Beata, M. Se, Ge Gavriouchov Sergeuei, M. Se, Nmr

151 Gaweł Damian, M. Se, Mu Góra Anna, M. Se, Pr Góra Monika, M. Se, Ge Grabińska Kariona, M. Se, Li Grabowska Dorota, M. Se, Li Grąziewicz Maria, M. Se, Mo Gromadka Robert, M. Se, Pr Grynberg Marcin, M. Se, Ge Grzybowska Ewa, M. Se, Ge Hałas Agnieszka, M. Se, Ge Janik Anna, M. Se, Li Janowicz Aleksandra, M. Se, Mi Joukov Igor, M. Se, Nmr Juszczuk Marek, M. Se, Mi Jurkowski Ireneusz, M. Se, PI Kacprzak Magdalena, M. Se, Ge Kaczanowski Szymon, M. Se, Inf Kamińska Joanna, M. Se, Ge Kazimierczak Beata, M. Se, Li Kierzek Andrzej, M. Se, Inf Konopińska Agata, M. Se, Ge Koprowski Piotr, M. Se, Mu Koter Marek, M. Se, PI Krysiak Cezary, M. Se, PI Kucharczyk Róża, M. Se, Pr Krzymowska Magdalena, M. Se, PI Lipska-Dwużnik Anna, M. Se, Pr Liwosz Aneta, M. Se, PI Maciejewska Agnieszka, M. Se, Mo Maliszewska-Tkaczyk Magdalena, M. Se, Mu Mieczkowski Piotr, M. Se, Mu Migdalski Andrzej, M. Se, Pr Mikołajczyk Monika, M. Se, PI Mikołajek-Zielińska Beata, M. Se, Nmr Milner Małgorzata, M. Se, Pr Musielak Małgorzata, M. Se, PI Nowosielska Anetta, M. Se, Mo Perlińska-Lenart Urszula, M. Se, Li Pluta Krzysztof, M. Se, Mi Podstolski Wojciech, M. Se, Pr Racki Waldemar, M. Se, Pr Smaczyńska-de Rooij Iwona, M. Se, Ge Sadowy Ewa, M. Se, Mi Sitkiewicz Izabela, M. Se, Mi Speina Elżbieta, M. Se, Mo

152 Szczegielniak Jadwiga, M. Se, PI Szymańska Małgorzata, M. Se, Li Talarczyk Andrzej, M. Se, PI Trojanek Joanna, M. Se, PI Tomaszewski Radosław, M. Se, Pr Wanke Małgorzata, M. Se, Li Waszkowska Ewa, M. Se, Mo Węgleńska Anna, M. Se, Mi Wójcik Anna, M. Se, Mo Zdanowski Konrad, M. Se, Bi Zielenkiewicz Urszula, M. Se, Mi

Holders of Ministry of Education Scholarships:

Malik Fateh Muhammad Avan, M. Se, PI Awotunde Olubunmi Sunday, M. Se, PI Vo Sy Hung, M. Se, Li

M. Se. Students:

Małgorzata Palczewska, Mi Monika Siedlecka, Nmr Magdalena Chełchacz, Pr Jakub Godlewski, Pr Agnieszka Borowska, Ge Celina Kaczor, Mi Piotr Kowalczyk, UW Michał Malewicz, UW Ewa Czernieć, Mi Małgorzata Jaworska, Mo Bernarda Kusztykiewicz, Mo Agnieszka Janisz, Mo Daniel Laubitz, Mo Paweł Kowalczyk, Pr Marcin Chouda, Li Ewelina Żak, Pr Marcin Górski, Pr Agnieszka Kucharska, Ge Alicja Jesionowska, Ge Lidia Polkowska, Pl Karolina Skorupińska, Li Elżbieta Dziedowiec, Mi Marcin Gołębiewski, Pr

153 Olga Bugajska, Mi Michał Skrzycki, Mi Bożena Bukowska, Mo Anna Wiśniewska, Mi Wojciech Dajewski, Mu Agnieszka Wiktorowska-Jezierska, Bi Małgorzata Mozoluk, Pl Anna Wójtowicz, Bi Anna Niżankowicz, Pl Katarzyna Onyszczuk, Pl Maksymilian Zienkiewicz, Mo Sławomir Krawczyk, Inf Magdalena Kowalczyk, Mi Dominika Krakowian, Mi Wojciech Augustyniak, Mo Katarzyna Lichota, Mo

154