ISSN 0077-4995 NATIONAL INSTITUTE OF GENETICS JAPAN ANNUAL REPORT No. 45 1994 Published by THE NATIONAL INSTITUTE OF GENETICS Mishima, Sizuoka-ken, Japan 1995 Annual Report of the National Institute of Genetics No. 45, 1994 ., •• • Published by The National Institute of Genetics, Japan 1995 CONTENTS General statement I Staff 3 Council and advisory committee 8 Association for propagation of the knowledge of genetics 10 Projects of research for 1994 11 Research activities in 1994 17 1. Molecular Genetics Molecular anatomy of RNA polymerase alpha subunit from Esche­ richia coil: functinal map of the amino-terminal assembly domain. KIMURA, M. and ISHIHAMA, A. .. ............ 17 Molecular anatomy of RNA polymerase alpha subunit: analysis of the carboxy-terminal activation domain by class-I factors and upstream enhancer elements. MURAKAMI, K., FUJITA, N., TAO, K., HEYDUK, T. and ISHIHANA, A. 18 Molecular anatomy of RNA polymerase alpha subunit: structure of the carboxy-terminal activation domain. NEGISHI, T., FUJITA, N., KYOGOKU, Y. and ISHIHAMA, A. 19 Promoter selectivity control of Escherichai coil RNA polymerase: intracellular levels of sigma subunits. JISHAGE, M., IWATA, A. and ISHIHAMA, A. .............................. .. 20 Promoter selectivity control of Escherichia coil RNA polymerase: functional comparison of sigma subunits. KUSANO, S., FUJITA, N. and ISHIHAMA, A. ... .. ..... .. .... ..... ... 21 Molecular anatomy of Escherichia coil RNA polymerase: modes of RNA polymerase-transcription factor interaction (progress in international collaborative studies). ISHIHAMA, A. 22 Gene cloning and characterization of RNA polymerase II subunits from the fission yeast Schizosaccharomyces pombe: cloning of the genes encoding small subunits. SAKURAI, H., SUZUKI, H. and ISHIHAMA, A. ................................ .. 24 Gene cloning and characterization of RNA polymerase II subunits from the fission yeast Schizosaccharomyces pombe: thermolabile ii RNA polymerase II with mutations in subunit 3. YASUI, K., AZUMA, Y., YAMAGISHI, M. and ISHIHAMA, A. .. 25 Gene cloning and characterization of RNA polymerase II subunits from the fission yeast Schizosaccharomyces pombe: structure and function of subunit 5. MIYAo, T., AZUMA, Y, SAKU­ RAI, H., YAMAGISHI, M. and ISHIHAMA, A. ... 0 0 •••• 0 • 26 Molecular anatomy of influenza virus RNA polymerase: analysis of subunit functions. KOBAYASHI, M., TOYODA, T., NAKADA, S., MlzuMoTo, Ko and ISHIHAMA, Ao 0 0 • 0 •• 0 0 0 0 •• 0 0 0 0 •• 26 Molecular anatomy of influenza virus RNA polymerase: analysis of subunit-subunit contact sites. ADYSHEV, D., TOYODA, T., IWATA, A., VEDA, S. and ISHIHAMA, A. 00 ••• 00 28 Molecular anatomy of influenza virus RNA polymerase: mapping of nucleotide-binding sites. ASANO, Y, MlzuMoTo, K., MARUYAMA, T. and ISHIHAMA, A. 0 •• 0 • 0 0 •••••••• 0 0 0 0 • 28 Molecular dissection of influenaza virus nucleoprotein (NP): iden- tification of RNA-binding region. KOBAYASHI, M., Toyo­ DA, T. and ISHIHAMA, Ao .. 0 0 0 0 • 0 •• 0 •• 0 ••••••••••• 0 0 0 29 Growth control of influenza virus by membrane protein (MI). YASUDA, J. and ISHIHAMA, A. .. 0 ••••••• 0 0 •••• 0 0 •• 0 0 0 • 29 Molecular dissection of mouse Mx1 protein with anti-influenza virus activity. TOYODA, T., ASANO, Y. and ISHIHAMA, A. 30 DNA replication/repair is impaired in a temperature-sensitive ubi­ quitin-activating enzyme mutant of mouse cells,without the accompanied enhanced mutation frequency-a possible in­ volvement of deoxyribonucleotide metabolism. AOKI, F., WATAYA, Y. and SENO, T. 0 ••••• 0. 0 ••••••••• 00 •• 00.0 •• 31 Perturbation of nucleolar dissolution in the G2-M transition in a mouse cell mutant with temperature-sensitive ubiquitin­ activating enzyme EI: possible involvement of HSP70 and proteasome. SUDHA To, SAMASHIMA, M., TSUJI, H., YAMAO, F. and SENO, T. .. 0000•0•00•0 ••••••••••• 0 0 0 0 32 Vbiquitin-activating Enzyme, El is phosphorylated in mammalian cells by the protein kinase CDC2. NAGAI, Y., KANEDA, S., SENO, To and YAMAO, F. 0 0 0 0 ••••••••••• 0 ••••••• 0 0 0 0 0 33 Screening for cDNAs encoding the ubiquitin-conjugating enzyme CONTENTS iii in fission yeast, Schizosaccharomyces pombe. OSAKA, F., SEINO, H., SENO, T. and YAMAO, F. 33 Regulation of G liS transition by the ubiquitin system in yeast Saccharomyces cerevisiae. KISHI, T., VI, M. and SENO, T. 34 Cell cycle regulation by the ubiquitin system in eukaryote. SEINO, H., YAGURA, T., YAMAO, F. and SENO, T. 35 Crystallographic studies of proteins involved in DNA repair. MORIKAWA, K . 36 Kinetic study on transcription using immobilized operons: branc- hed pathway in early elongation. KUBORI, T. and SHIMA- MOTO, N. 38 Single-molecule dynamics of transcription using immobilized ope­ rons: sliding of P. putida CamR protein on DNA. KABATA, H. and SHIMAMOTO, N . 38 Functional and structural study of E. coil single-stranded DNA binding protein. NAGAI, H. and SHIMAMOTO, N. 39 In vitro transcriptional analysis of the Bacillus subtilis sporulation gene. FUJITA, M. and SADAIE, Y. 40 II. Microbial Genetics Recognition of the minus strand DNA replication origin of the filamentous phage by E. coli RNA polymerase. GUAN, Z. W., HIGASHITANI, N., HIGASHITANI, A. and HORIUCHI, K. 41 The psv gene coding for penicillin-binding protein 7 of Escherichia coli. HARA, H., ABE, N., NAKAKOHJI, M. NISHIMURA, Y. and HORIUCHI, K. ................................ .. 43 A cell division inhibitor SulA of Escherichia coli directly interacts with FtsZ through GTP hydrolysis. HIGASHITANI, A. and HORIUCHI, K. .................................... .. 43 Isolation and characterization of mutations that suppress the effect of a cell-division inhibitor SulA in Escherichia coli. HIGASHI- TANI, A. and HORIUCHI, K. .. 45 Molecular mechanisms that determine the timing of cell division in Escherichia coli K12. NISHIMURA, A. 46 A new gene which couples cell growth and cell division in Escheri- chia coli K-12. VKAI, H., WATANABE, N. and NISHIMURA, iv A . 47 The role of the secA protein of Bacillus subtillis in sporulation. SADAIE, Y. and FUJITA, M. 48 III. Mammalian Genetics Genetical analysis of rim2 coat color mutaion. SAGAI, T., W AKANA, S., MORIWAKI, K. and SHIROISHI, T. .. 49 Meiotic recombination at the Lmp2 hotspot in the mouse major histocompatibility complex is tolerant of substitutional base mismatches. YOSHINO, M., SAGAI, T and SSHIROISHI, T. 50 Genetic analysis of mouse mutants affecting limb pattern formation. MASUYA, H., SAGAI, T, MORIWAKI, K. and SHIRaIsHI, T. 52 Linkage analysis of the skeleton mouse mutant, Tail short (Ts). SHIROISHI, T, UCHIDA, K., KOOPMAN, P., MlTA, A., WAKANA, S. and MORIWAKI, K. 53 Gene mapping toward positional cloning of the Rim3 gene. MASUYA, H., WAKANA, S., SAGAI, T., MORIWAKI, K. and SHIRaIsHI, T. 54 Meitotic recombination on DNase I-hypersensitive sites in the mouse major histocompatibility complex. MIZUNO, K-I., SAGAI, T., MORIWAKI, K. and SHIROISHI, T. 54 IV. Developmental Genetics Sexual differentiation potency of multi-potent stem cells in hydra. NISHIMIYA-FuJISAWA, C. and SUGIYAMA, T . 56 Hydra regeneration from recombined ectodermal and endodermal tissue. I. Cell rearrangement. KISHIMOTa, Y, MURATE, M. and SUGIYAMA, T. 57 Hydra regeneration from recombined ectodermal and endodermal tissue. II. Electron microscopic study. MURATE, M., KISHI­ MOTa, Y., SUGIYAMA, T., TAKAHASHI-IWANAGA, H. and IWANAGA, T . 58 Isolation of cytoskeletal molecules in hydra by epitope selection. HATTA, M., SAKAGUCHI, M., KOBAYAKAWA, Y., KOIZUMI, O. and SUGIYAMA, T. 59 CONTENTS v Disappearance and reappearance of mesoglea in hydra tissue during regeneration after head removal. SHIMIZU, H. and SARRAS, M. J. Jr. 000000000000000000.000 ••••••••••••••• 60 Hydra peptide project. I. Isolation,structure determination and chemical synthesis. MUNEOKA, Yo, TAKAHASHI, T., FUJI­ SAWA, T, SHIMIZU, H., HATTA, M., SUGIYAMA, T., Kor­ ZUMI, 0., BOSCH, T., DEHARO, M., LOHMANN, J., DAVID, C. and BODE, H. 00.0 ••••••••••••••• 00.0000000000000000 62 Hydra peptide project. II. Differential display-PCR assay for signal molecule activity. FUJISAWA, T., SHIMIZU, H., HATTA, M., SUGIYAMA, T., BOSCH, T., DEHARO, M., LOHMANN, J., DAVID, C., TAKAHASHI, T., MUNEOKA, Y., KorZUMI, o. and BODE, H. .000.000 •••••••••••••••• 00 •• 00000000000000 63 Hydra peptide project. III. Biological function assay. FUJISAWA, T, SHIMIZU, H., HATTA, M., SUGIYAMA, T, KOIZUMI, 0., BODE, H, BOSCH, T., DEHARO, M., LOHMANN, J., DAVID, C., TAKAHASHI, T. and MUNEOKA, Y. 64 Intermittent expression of BmFTZ-Fl, a member of the nuclear hormone receptor superfamily during development of the silk­ worm Bombyx mori. SUN, G.-C., HIROSE, S. and VEDA, H. 65 Mediators of activation of jushi tarazu gene transcription by BmFTZ-Fl. LI, F.-Q., VEDA, H. and HIROSE, S. 0 66 Sequences of two cDNAs encoding silkworm homologues of Dro­ sophila melanogaster squid gene. LI, F.-Q., SUN, G.-c., VEDA, H. and HIROSE, S. 0 • 0 ••• 00000000 66 The Drosophila nuclear receptors FTZ-Fla and FTZ-Fl,B compete as monomers for binding to a site in the jushi tarazu gene. OHNO, C. K., VEDA, H. and PETKOVICHI, M. .. 0 0 0 0 0 0 0 0 67 The Bombyx silkworm is a semslparous insect with a long life in the post-reproductive period. MURAKAMI, A. 0 0 •••••••••••• 68 Homoeostasis in the process of growth and senility in the Bombyx silkworm: the relationship between Larval growth period and temperature. MURAKAMI, A. 0 • 0 0 0 0 •••••••••••••••••• 69 Homoeostasis in growth and senility in the Bombyx silkworm: the relationship between adult lifespan and body-weight. MURA-