DOCSLIB.ORG

The Importance of Mutation, Then and Now: Studies with Yeast Cytochrome C

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Importance of Mutation, Then and Now: Studies with Yeast Cytochrome C 2 F. Sherman / Mutation Research 589 (2005) 1–16 Mutation Research 589 (2005) 1–16 Contents 1. My first encounter with yeast research . .................................................. 2 The importance of mutation, then and now: 2. Identification of CYC1 ............................................................... 3 Mutation Research 589 (2005) 1–16 ☆ 3. Importance of cytochrome c ........................................................... 4 studies with yeast cytochrome c www.elsevier.com/locate/reviewsmr Community address: www.elsevier.com/locate/mutres Fred Sherman * Reflections in Mutation Research 4. CYC1 encodes iso-1-cytochrome c ....................................................... 4 5. Isolation and characterization of cyc1 mutants. .............................................. 5 Department ofThe Biochemistry importance and Biophysics, of University mutation, of Rochester then Medical andSchool, now:Box 712, Rochester, studies NY 14642, USA Accepted 26 July 2004 with yeast cytochrome c$ 6. Generating altered genes and proteins, then and now .......................................... 6 Available online 23 September 2004 7. Deducing DNA sequences from protein sequences. .......................................... 8 Fred Sherman* 8. ‘‘Site-directed’’ mutagenesis in early times . .............................................. 10 Department of Biochemistry and Biophysics, University of Rochester Medical School, Box 712, Rochester, NY 14642, USA 9. Cloning CYC1 and transcriptional regulation. .............................................. 10 Accepted 26 July 2004 Available online 23 September 2004 10. Subsequent and current studies . ...................................................... 11 11. A final reflection ................................................................... 14 Abstract Acknowledgements . ................................................................... 14 The development of a genetic system based on the CYC1 gene was initiated over 40 years ago, primarily because of the anticipated ease of sequencing of the corresponding encoded protein, iso-1-cytochrome c from Saccharomyces cerevisiae. The References ........................................................................... 14 success of the iso-cytochrome c system was dependent on the early development of methods for detecting and selecting cyc1 defective mutants and CYC1 functional revertants, and of methods for fine-structure genetic mapping using deletions and single- site mutations. The nonsense codons TAA and TAG, and the initiation codon ATG, were determined from the amino acid 1. My first encounter with yeast research emphasized to me that the existence of bacteriophages alterations of iso-1-cytochromes c from intragenic revertants; this represented the first assignments of such codons in a eukaryotic organism. The types of desired sequences were expanded by selecting recombinants from cyc1 cyc1 nonfunctional was one of the major advantages for using E. coli, and mutants or CYC1 CYC1 functional mutants, permitting the early determination of the rules of translation,� which differed from I began using yeast, Saccharomyces cerevisiae, as it was suggested that I find phages acting similarly on � those of prokaryotes by use of the most 50 AUG codon for initiation of translation. The sequence of 44 base pairs of CYC1 was an experimental organism 50 years ago, when I first yeast. My immediate response was ‘‘where do I find determined with altered iso-1-cytochromes c from revertants of frameshift and initiation mutants, allowing the early cloning of the joined the biophysics graduate program at Berkeley in them?’’ It was explained to me that the natural habitat gene. A method was developed for transforming yeast directly with synthetic oligonucleotides, resulting in the convenient 1954. Soon after my arrival, the director of the program of yeast was on the surface of fruits and that tropical production of CYC1 mutants with defined sequences. At this point in time, Sherman and colleagues have published approximately discussed a possible research project involving yeast. birds eat fruit. Therefore, a likely source of yeast 240 papers on or using the iso-cytochrome c system, dealing with such diverse topics as translation, informational suppressors, In this regard, one of the main interests of biophysicists phages was tropical bird feces. At that moment a transcription and transcription termination, recombination, ectopic recombination, mutagen specificity, regulation by Ty1 at that time was target theory and the effects of ionizing thought flashed through my mind, ‘‘I am going to be elements, evolution of duplicated chromosomal segments, structure–function relationships of cytochrome c, protein stability radiation. This was a time when physicists overly sent to the jungles of the Amazon or the Congo and degradation, biosynthesis and mitochondrial import of cytochrome c, mitochondrial proteases, co- and post-translational interpreted survival curves and before DNA repair was to collect tropical bird feces.’’ ‘‘Where do I go to modifications, and mRNA degradation. Current work on degradation of proteins in mitochondria, on degradation of mRNA in the considered. Carl C. Lindegren had already described collect tropical bird feces?’’ I asked. The answer was nucleus, and on N-terminal acetylation stems from properties of CYC1 mutants isolated in early screens more than a decade ago. the heterothallic life cycle, and Robert K. Mortimer logical but disappointing, ‘‘The local San Francisco # 2004 Elsevier B.V. All rights reserved. was busy making a polyploid series of yeast, haploid Fleishhacker Zoo across the Bay Bridge.’’ My next Keywords: Cytochrome c; Nonsense codons; Inititation codon; Protein stability and degradation; N-terminal acetylation; mRNA degradation through hexaploid, a series which appeared to be useful question was ‘‘how do I get inside the cages?’’ I was for exploring target theory [1]. I was informed that reminded that I am now a graduate student and have to any new fundamental information on yeast would be be resourceful and independent. of considerable importance, since it was the up-and- I called the Zoo and asked to speak to the Director, $ This article is part of the Reflection in Mutation Research series. To suggest topics and authors for Reflection, readers should contact the series editors G.R. Hoffmann ([email protected]) or D.G. MacPhee ([email protected]). coming microorganism of the future. a Ph.D. in zoology. I explained to the Director that I * Tel.: +1 585 275 6647; fax: +1 585 275 6007. This was the time when Escherichia coli was was a graduate student at the University of Calfornia, E-mail address: [email protected]. clearly the model system for molecular biology. It was and that I would like to collect fresh tropical bird 1383-5742/$ – see front matter # 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.mrrev.2004.07.001 200 ELSEVIER REFLECTIONS IN MUTATION RESEARCH: 1999 – 2019 201 F. Sherman / Mutation Research 589 (2005) 1–16 3 4 F. Sherman / Mutation Research 589 (2005) 1–16 feces. There was pause, then a hesitant response, ‘‘Oh, chronize cells by heat shock. I attempted to deduce the the RAD7 and OSM1 loci as well as the CYC1 locus encompassing the heme group was already sequenced you mean bird shit—yeah, that’s OK.’’ number of genetic determinants in a cell by [5–7]. Some 5 years later, a second deletion, cyc1-237, [13], and entire cytochromes c from various species The next day I arrived at the Zoo fully equipped mathematically modeling the kinetics of rÀ induction with the same seemingly pleiotropic set of phenotypes were being sequenced in several laboratories. Thus, with vials, spatulas, a notebook and my roommate, a after growth at elevated temperatures. After receiving was unexpectedly uncovered among meiotic segre- investigating cytochrome c appeared to be an ideal pre-law student who was responsible for recording the my degree, I investigated recombination in yeast as a gants. The origin of this type of deletion was not project, especially because yeast had the advantage of species of the birds and the time of collection of the Post-doctoral Fellow at Seattle with the late Herschell understood until Liebman et al. [8] noted that certain being a eukaryotic microorganism with a well-defined specimens. We were both adorned with official- Roman. Subsequently, I entered a second post- laboratory strains spontaneously gave rise to high genetic system. The advantages of using yeast looking white lab coats. After entering an aviary, a doctoral position in the laboratory of the late Boris frequencies of deletions encompassing the CYC1, cytochrome c became even more evident a few years crowd gathered in front of the cage and it soon became Ephrussi at Gif-sur-Yvette, France, where I initiated a OSM1 and RAD7 genes and that the deletions were later when Narita et al. [14] announced the complete clear to them what our purpose was. Apparently, we research program in 1960 that led to the discovery of flanked by Ty1 elements [6,9]. Thus, cyc1-1, the first amino acid sequence of iso-1-cytochrome c. were more interesting than the animals. As soon as a the CYC1 gene. cytochrome c deficient mutant, was uncovered because bird defecated, several members of the crowd pointed of the rare occurrence of a spontaneous deletion in a and shouted ‘‘There’s one, there’s one.’’ strain containing an unrelated pet mutation. 4. CYC1 encodes iso-1-cytochrome c After returning to the University and refrigerating 2.
Load more

Recommended publications
  • Dual Mutations Reveal Interactions Between Components of Oxidative Phosphorylation in Kluyveromyces Lactis
    Copyright 2001 by the Genetics Society of America Dual Mutations Reveal Interactions Between Components of Oxidative Phosphorylation in Kluyveromyces lactis G. D. Clark-Walker and X. J. Chen Molecular Genetics and Evolution Group, Research School of Biological Sciences, The Australian National University, Canberra, ACT, 2601, Australia Manuscript received April 10, 2001 Accepted for publication August 3, 2001 ABSTRACT Loss of mtDNA or mitochondrial protein synthesis cannot be tolerated by wild-type Kluyveromyces lactis. The mitochondrial function responsible for ␳0-lethality has been identified by disruption of nuclear genes encoding electron transport and F0-ATP synthase components of oxidative phosphorylation. Sporulation of diploid strains heterozygous for disruptions in genes for the two components of oxidative phosphoryla- tion results in the formation of nonviable spores inferred to contain both disruptions. Lethality of spores is thought to result from absence of a transmembrane potential, ⌬⌿, across the mitochondrial inner membrane due to lack of proton pumping by the electron transport chain or reversal of F1F0-ATP synthase. Synergistic lethality, caused by disruption of nuclear genes, or ␳0-lethality can be suppressed by the atp2.1 ␤ mutation in the -subunit of F1-ATPase. Suppression is viewed as occurring by an increased hydrolysis of ATP by mutant F1, allowing sufficient electrogenic exchange by the translocase of ADP in the matrix for ATP in the cytosol to maintain ⌬⌿. In addition, lethality of haploid strains with a disruption of AAC encoding the ADP/ATP translocase can be suppressed by atp2.1. In this case suppression is considered to occur by mutant F1 acting in the forward direction to partially uncouple ATP production, thereby stimulating respiration and relieving detrimental hyperpolarization of the inner membrane.
    [Show full text]
  • Attachment 1
    Appendix 1 Chemico-Biological Interactions 301 (2019) 2–5 Contents lists available at ScienceDirect Chemico-Biological Interactions journal homepage: www.elsevier.com/locate/chembioint An examination of the linear no-threshold hypothesis of cancer risk T assessment: Introduction to a series of reviews documenting the lack of biological plausibility of LNT R. Goldena,*, J. Busb, E. Calabresec a ToxLogic, Gaithersburg, MD, USA b Exponent, Midland, MI, USA c University of Massachusetts, Amherst, MA, USA The linear no-threshold (LNT) single-hit dose response model for evolution and the prominence of co-author Gilbert Lewis, who would be mutagenicity and carcinogenicity has dominated the field of regulatory nominated for the Nobel Prize some 42 times, this idea generated much risk assessment of carcinogenic agents since 1956 for radiation [8] and heat but little light. This hypothesis was soon found to be unable to 1977 for chemicals [11]. The fundamental biological assumptions upon account for spontaneous mutation rates, underestimating such events which the LNT model relied at its early adoption at best reflected a by a factor of greater than 1000-fold [19]. primitive understanding of key biological processes controlling muta- Despite this rather inauspicious start for the LNT model, Muller tion and development of cancer. However, breakthrough advancements would rescue it from obscurity, giving it vast public health and medical contributed by modern molecular biology over the last several decades implications, even proclaiming it a scientific principle by calling it the have provided experimental tools and evidence challenging the LNT Proportionality Rule [20]. While initially conceived as a driving force model for use in risk assessment of radiation or chemicals.
    [Show full text]
  • Barbara Mcclintock's World
    Barbara McClintock’s World Timeline adapted from Dolan DNA Learning Center exhibition 1902-1908 Barbara McClintock is born in Hartford, Connecticut, the third of four children of Sarah and Thomas Henry McClintock, a physician. She spends periods of her childhood in Massachusetts with her paternal aunt and uncle. Barbara at about age five. This prim and proper picture betrays the fact that she was, in fact, a self-reliant tomboy. Barbara’s individualism and self-sufficiency was apparent even in infancy. When Barbara was four months old, her parents changed her birth name, Eleanor, which they considered too delicate and feminine for such a rugged child. In grade school, Barbara persuaded her mother to have matching bloomers (shorts) made for her dresses – so she could more easily join her brother Tom in tree climbing, baseball, volleyball, My father tells me that at the and football. age of five I asked for a set of tools. He My mother used to did not get me the tools that you get for an adult; he put a pillow on the floor and give got me tools that would fit in my hands, and I didn’t me one toy and just leave me there. think they were adequate. Though I didn’t want to tell She said I didn’t cry, didn’t call for him that, they were not the tools I wanted. I wanted anything. real tools not tools for children. 1908-1918 McClintock’s family moves to Brooklyn in 1908, where she attends elementary and secondary school. In 1918, she graduates one semester early from Erasmus Hall High School in Brooklyn.
    [Show full text]
  • From the President's Desk
    JAN/FEB 2006 From the President’s desk: 2006, the 75th anniversary of the Genetics Society of America, will be marked by a number of initiatives to reinvigorate the Society’s mission of promoting research and education in genetics. A highlight was the recently held GSA sponsored conference, “Genetic Analysis: From Model Organisms to Human Biology” in San Diego from January 5-7. This conference emphasized the importance of model organism research by illustrating the crucial contributions to human biology resulting from discoveries in these organisms. The National Institutes of Health (NIH) supported this conference both financially and by participation of key NIH administrators, including Jeremy M. Berg, director of the National Institute of General Medical Sciences. In addition to the superb science talks by international leaders the MOHB conference showcased other important and new GSA initiatives including education, public policy advocacy, graduate student support and recognition of outstanding model organism geneticists. Robin Wright, Education Committee chair, led a round table discussion on undergraduate education and the Joint Steering Committee for Public Policy and the Congressional Liaison Committee sponsored a session on science advocacy and public policy. There was a mentor lunch to support graduate students and postdocs in the next steps of their careers, and the three GSA medals were presented during the banquet, with Victor Ambros receiving the GSA Medal, Fred Sherman the Beadle Award, and Masatoshi Nei the Morgan Award. (For research highlights at the meeting, see pages 6 and 7 of this issue.) The 75th anniversary will also usher in changes to our society’s journal, GENETICS.
    [Show full text]
  • Fred Sherman: a Pioneer in Genetics
    CLASS NOTES TRIBUTE Fred Sherman: A Pioneer in Genetics Fred Sherman, a pioneer in genetics and long hours of laboratory work, large doses mance of Indian dance (dance was a major molecular biology, was a member of the of Fred’s zany humor, and after-hours interest and activity of Fred’s), his thoughts Rochester faculty for 52 years, from 1961 sampling of the local night-life. It provided and conversation quickly returned to yeast. until his death last September. The breadth an introduction to yeast for many scientists His encyclopedic memory of 50 years of of his scientific contributions over the span who went on to become leaders in modern yeast genetics included much information of years that saw the development of that was never published and is now, modern molecular biology is simply sadly, lost. Fortunately, his engagement breathtaking. Fred’s early scientific with science was tempered by humor studies focused on the gene encoding that found expression both in a staple the protein cytochrome c in baker’s of often-repeated jokes for every occa- yeast, establishing this as a power- sion and in carefully crafted comedic ful system that allowed him to make remarks that he proffered in the guise fundamental contributions to the initial of questions and comments at scientific deciphering of the genetic code. His seminars. He semi-seriously referred to determination of the DNA sequence himself as the world’s expert in yeast of the gene encoding cytochrome c, genetic nomenclature, and once tried one of the first eukaryotic genes to be to win a dispute with an editor about sequenced, served as the basis for the the naming of a particular yeast gene first cloning of a gene from a eukary- by announcing that he had tattooed otic organism.
    [Show full text]
  • Molecular and Cellular Biology
    MOLECULAR AND CELLULAR BIOLOGY VOLUME 3 * NUMBER 1 o JANUARY 1983 Aaron J. Shatkin, Editor-in-Chief (1985) Roche Instituite of Molecular Biology Nutlex, N.J. Harvey F. Lodish, Editor (1986) Louis Siminovitch, Editor (1985) Massachuisetts Institute of Technology Hospital for Sick Children Cambridge Toronto, Canadal David J. L. Luck, Editor (1987) Paul S. Sypherd, Editor (1985) Rockefeller Unis'ersity University of California Ness York, N.Y. Irvine EDITORIAL BOARD Renato Baserga (1985) Ira Herskowitz (1984) Daniel B. Rifkin (1985) Alan Bernstein (1984) Larry Kedes (1985) Robert G. Roeder (1985) J. Michael Bishop (1984) Marilyn Kozak (1985) James E. Rothman (1984) Joan Brugge (1985) Elias Lazarides (1985) David Sabatini (1985) Breck Byers (1985) John B. Little (1985) Phillip A. Sharp (1985) John A. Carbon (1984) William F. Loomis, Jr. (1985) Fred Sherman (1985) Lawrence A. Chasin (1985) Paul T. Magee (1985) Pamela Stanley (1985) Nam-Hai Chua (1985) Robert L. Metzenberg (1985) Joan A. Steitz (1985) Terrance G. Cooper (1984) Robert K. Mortimer (1985) James L. Van Etten (1985) James E. Darnell, Jr. (1985) Harvey L. Ozer (1985) Jonathan R. Warner (1984) Gary Felsenfeld (1985) Mary Lou Pardue (1985) Robert A. Weinberg (1984) Norton B. Gilula (1985) Mark Pearson (1985) I. Bernard Weinstein (1985) James E. Haber (1984) Jeremy Pickett-Heaps (1985) Harold Weintraub (1985) Benjamin Hall (1985) Robert E. Pollack (1985) Reed B. Wickner (1985) Ari Helenius (1984) Keith R. Porter (1985) Leslie Wilson (1985) Susan A. Henry (1985) John R. Pringle (1985) Edward Ziff (1985) Helen R. Whiteley, Chairman, Publications Board Walter G. Peter III, Director, Puiblications Linda M.
    [Show full text]
  • Original Article Bioinformatic Analysis of Gene Expression Profile in Prostate Epithelial Cells Exposed to Low-Dose Cadmium
    Int J Clin Exp Med 2018;11(3):1669-1678 www.ijcem.com /ISSN:1940-5901/IJCEM0062792 Original Article Bioinformatic analysis of gene expression profile in prostate epithelial cells exposed to low-dose cadmium Qiling Liu1,2*, Rongqiang Zhang2*, Xiang Wang1, Peili Wang2, Xiaomei Ren2, Na Sun2, Xiangwen Li2, Xinhui Li2, Chunxu Hai1 1Department of Toxicology, The Ministry of Education Key Lab of Hazard Assessment and Control in Special Op- erational Environment, Shaanxi Provincial Key Lab of Free Radical Biology and Medicine, School of Public Health, Medical University of The Air Force, Xi’an, Shaanxi 710032, China; 2Department of Epidemic and Health Statis- tics, The College of Public Health for The Shaanxi University of Chinese Medicine, Shaanxi 712046, China. *Equal contributors. Received July 25, 2017; Accepted February 5, 2018; Epub March 15, 2018; Published March 30, 2018 Abstract: Objective: This study was to identify key genes and biological pathways involved in responses of prostate epithelial cells after low-dose Cd exposure by using bioinformatic analysis. Methods: The gene chip data of prostate epithelial cells after low-dose Cd exposure were collected from public databases Gene Expression Omnibus. After identification of differentially expressed genes (DEGs), data were input into Qlucore Omics Explorer, Network Analyst, String, and Genclip for further analysis of gene expression profiles, protein-protein interactions (PPI) and protein- chemicals interactions, and critical molecular pathways. Results: A total of 384 DEGs were identified in Cd treated group compared with control group. The number of DEGs gradually decreased over time, with the largest number at 0 h. Furthermore, NDUFB5 (A, S), CYC1, UQCRB, ETFA (B), SNRPD2, and LSM3 (5, 6) were the hub proteins in the PPI network.
    [Show full text]
  • Moore Noller
    2002 Ada Doisy Lectures Ada Doisy Lecturers 2003 in BIOCHEMISTRY Sponsored by the Department of Biochemistry • University of Illinois at Urbana-Champaign Dr. Peter B. 1970-71 Charles Huggins* and Elwood V. Jensen A76 1972-73 Paul Berg* and Walter Gilbert* Moore 1973-74 Saul Roseman and Bruce Ames Department of Molecular carbonyl Biophysics & Biochemistry Phe 1974-75 Arthur Kornberg* and Osamu Hayaishi Yale University C75 1976-77 Luis F. Leloir* New Haven, Connecticutt 1977-78 Albert L. Lehninger and Efraim Racker 2' OH attacking 1978-79 Donald D. Brown and Herbert Boyer amino N3 Tyr 1979-80 Charles Yanofsky A76 4:00 p.m. A2486 1980-81 Leroy E. Hood Thursday, May 1, 2003 (2491) 1983-84 Joseph L. Goldstein* and Michael S. Brown* Medical Sciences Auditorium 1984-85 Joan Steitz and Phillip Sharp* Structure and Function in 1985-86 Stephen J. Benkovic and Jeremy R. Knowles the Large Ribosomal Subunit 1986-87 Tom Maniatis and Mark Ptashne 1988-89 J. Michael Bishop* and Harold E. Varmus* 1989-90 Kurt Wüthrich Dr. Harry F. 1990-91 Edmond H. Fischer* and Edwin G. Krebs* 1993-94 Bert W. O’Malley Noller 1994-95 Earl W. Davie and John W. Suttie Director, Center for Molecular Biology of RNA 1995-96 Richard J. Roberts* University of California, Santa Cruz 1996-97 Ronald M. Evans Santa Cruz, California 1998-99 Elizabeth H. Blackburn 1999-2000 Carl R. Woese and Norman R. Pace 2000-01 Willem P. C. Stemmer and Ronald W. Davis 2001-02 Janos K. Lanyi and Sir John E. Walker* 12:00 noon 2002-03 Peter B.
    [Show full text]
  • Fred Sherman an Introduction to the Genetics and Molecular
    Fred Sherman An Introduction to the Genetics and Molecular Biology of the Yeast Saccharomyces cerevisiae. (2001) http://dbb.urmc.rochester.edu/labs/sherman_f/yeast/index.html Table of Contents • 1 Yeast as a Model Eukaryote • 11 Manipulating the Genome In • 2 Information on Yeast Vitro with Plasmids • 3 Yeast Strains 11.1 Cloning by Complementation • 4 Growth and Life Cycles 11.2 Mutagenesis In Vitro • 5 The Yeast Genome 11.3 Two-step Gene Replacement • 6 Genetic Nomenclature 11.4 Gene Disruption and One-step 6.1 Chromosomal Genes Gene Replacement 6.2 Mitochondrial Gene 11.5 Plasmid Shuffle 6.3 Non-Mendelian Determinants 11.6 Recovering mutant alleles • 7 Genetic Analyses • 12 Interactions of Genes 7.1 Overviews with Examples 12.1 Heterozygosity and Dominant- 7.2 Tetrad Analysis negative Mutations 7.3 Non-Mendelian Inheritance 12.2 Intragenic Complementation • 8 Transformation 12.3 Nonallelic Non- 8.1 Yeast Vector and DNA complementation Fragments 12.4 Suppressors 8.2 Synthetic Oligonucleotides 12.5 Synthetic Enhancement and 8.3 Mitochondrial Transformation Epistatic Relationships • 9 Yeast Vectors • 13 Genomic Analysis 9.1 YIp Vectors • 14 Analyses with Yeast Systems 9.2 YEp Vectors 14.1 Two-hybrid Systems 9.3 YCp Vectors 14.2 Yeast Artificial Chromosomes • 10 Genes Important for Genetic (YACs) Studies 14.3 Expression of Heterologous 10.1 URA3 and LYS2 Protein in Yeast 10.2 ADE1 and ADE2 • Key Words 10.3 GAL1 Promoter • Bibliography 10.4 lacZ and Other Reporters 1 1 Yeast is a Model Eukaryote mutations can be conveniently isolated and manifested in haploid strains, and This chapter deals only with the yeast S.
    [Show full text]
  • MCDB 5220 Methods and Logics April 21 2015 Marcelo Bassalo
    Cracking the Genetic Code MCDB 5220 Methods and Logics April 21 2015 Marcelo Bassalo The DNA Saga… so far Important contributions for cracking the genetic code: • The “transforming principle” (1928) Frederick Griffith The DNA Saga… so far Important contributions for cracking the genetic code: • The “transforming principle” (1928) • The nature of the transforming principle: DNA (1944 - 1952) Oswald Avery Alfred Hershey Martha Chase The DNA Saga… so far Important contributions for cracking the genetic code: • The “transforming principle” (1928) • The nature of the transforming principle: DNA (1944 - 1952) • X-ray diffraction and the structure of proteins (1951) Linus Carl Pauling The DNA Saga… so far Important contributions for cracking the genetic code: • The “transforming principle” (1928) • The nature of the transforming principle: DNA (1944 - 1952) • X-ray diffraction and the structure of proteins (1951) • The structure of DNA (1953) James Watson and Francis Crick The DNA Saga… so far Important contributions for cracking the genetic code: • The “transforming principle” (1928) • The nature of the transforming principle: DNA (1944 - 1952) • X-ray diffraction and the structure of proteins (1951) • The structure of DNA (1953) How is DNA (4 nucleotides) the genetic material while proteins (20 amino acids) are the building blocks? ? DNA Protein ? The Coding Craze ? DNA Protein What was already known? • DNA resides inside the nucleus - DNA is not the carrier • Protein synthesis occur in the cytoplasm through ribosomes {• Only RNA is associated with ribosomes (no DNA) - rRNA is not the carrier { • Ribosomal RNA (rRNA) was a homogeneous population The “messenger RNA” hypothesis François Jacob Jacques Monod The Coding Craze ? DNA RNA Protein RNA Tie Club Table from Wikipedia The Coding Craze Who won the race Marshall Nirenberg J.
    [Show full text]
  • Molecular and Biochemical Characterisation of the Electron Transport Chain of Plasmodium Falciparum
    Molecular and biochemical characterisation of the electron transport chain of Plasmodium falciparum Thomas ANTOINE Ph.D. February 2012 Molecular and biochemical characterisation of the electron transport chain of Plasmodium falciparum Thesis is submitted in accordance with the requirements of the University of Liverpool for the degree of Doctor of Philosophy By Thomas ANTOINE February 2012 Declaration This thesis is the result of my own work. The material contained in the thesis has not been presented, nor is currently being presented, either wholly or as a part, for any other degree or other qualification. The research work was carried out in the Liverpool School of Tropical Medicine, University of Liverpool, United Kingdom. ……………………………………….. Thomas ANTOINE (2012) i Acknowledgments First of all, I would like to express my gratitude to my supervisors Pr. Steve Ward, Dr. Giancarlo Biagini and Dr. Nick Fisher for their supervision, continued support and constant encouragement throughout my PhD. A special thank goes to the InterMal PhD program and the Marie Curie Fellowship for providing the generous funding which allowed me to undertake this research, but also for giving me the opportunity to attend conferences and workshops as well as meet so many interesting people. I am grateful to Prof. Alister Craig and Prof. Sylke Muller for accepting to be the two members of my oral defense committee. The members of the Ward's research group have contributed immensely to my personal and professional time at Liverpool. I am very grateful to Ashley Warman for insightful comments in my work, for reviewing my thesis and providing valuable feedback as well as for many motivating discussions.
    [Show full text]
  • Identification of a Region of Homozygous Deletion on 8P22–23.1 in Medulloblastoma
    Oncogene (2002) 21, 1461 ± 1468 ã 2002 Nature Publishing Group All rights reserved 0950 ± 9232/02 $25.00 www.nature.com/onc Identi®cation of a region of homozygous deletion on 8p22 ± 23.1 in medulloblastoma Xiao-lu Yin1, Jesse Chung-sean Pang1 and Ho-keung Ng*,1 1Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China To identify critical tumor suppressor loci that are radiotherapy and chemotherapy, have greatly improved associated with the development of medulloblastoma, the outcomes of patients. In the past 30 years, the 5- we performed a comprehensive genome-wide allelotype year survival rate has increased from 10 to about 50%. analysis in a series of 12 medulloblastomas. Non-random However, long-term survival in children with advanced allelic imbalances were identi®ed on chromosomes 7q disease is only about 30% (Giangaspero et al., 2000; (58.3%), 8p (66.7%), 16q (58.3%), 17p (58.3%) and Heideman et al., 1997). Further enhancement of 17q (66.7%). Comparative genomic hybridization analy- survival will rely on a better understanding of the sis con®rmed that allelic imbalances on 8p, 16q and 17p biology of this malignant disease to improve current were due to loss of genetic materials. Finer deletion treatments or to develop novel therapy. mapping in an expanded series of 23 medulloblastomas Non-random loss of genetic material from chromo- localized the common deletion region on 8p to an interval somal loci is a common feature in the development of of 18.14 cM on 8p22 ± 23.2.
    [Show full text]