Abstract

The long-term objective of this project is to understand the molecular mechanisms that are responsible for induced mutagenesis in bakers' yeast, Saccharomyces cerevisiae. Since yeast is a eucaryote, such information is likely to prove of value to several health-related problems, specifically in understanding certain inherited diseases and as a theoretical basis for the evaluation of mutagenic and carcinogenic hazards.
The specific aims for the proposed funding period are of two kinds. First, the capability of yeast cells to carry out translesion synthesis will be critically evaluated, and the genetic and physiological conditions required examined. At the same time, the resulting spectrum of nucleotide sequence changes will be determined, the overall error-rate estimated, and the proportion of targeted and untargeted events established.
This aim will be achieved by constructing an M13 based yeast shuttle vector into which synthetic oligonucleotides containing a single-defined premutagenic lesion will be cloned. The second set of specific aims are to study the structure and function of the yeast genes concerned with induced mutagenesis, in particular the REV3 and newly isolated REV7 genes. Plasmids carrying these loci will be isolated by complementation of defective mutants, and the presence of the structural gene on the plasmid verified by integration and genetic mapping. After localization of the gene by subcloning and transcript mapping, the locus will be sequenced. LacZ fusions will be used to isolate antibodies to the gene product, and the latter isolated by immunoprecipitation.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM021858-12
Application #
3270741
Study Section
Radiation Study Section (RAD)
Project Start
1978-02-01
Project End
1991-01-31
Budget Start
1986-02-01
Budget End
1987-01-31
Support Year
12
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Rochester
Department
Type
Schools of Medicine
DUNS #
208469486
City
Rochester
State
NY
Country
United States
Zip Code
14627

  Publications

Li, Ziqiang; Zhang, Hong; McManus, Terrence P et al. (2002) hREV3 is essential for error-prone translesion synthesis past UV or benzo[a]pyrene diol epoxide-induced DNA lesions in human fibroblasts. Mutat Res 510:71-80
Lawrence, C W; Maher, V M (2001) Eukaryotic mutagenesis and translesion replication dependent on DNA polymerase zeta and Rev1 protein. Biochem Soc Trans 29:187-91
Lawrence, C W; Maher, V M (2001) Mutagenesis in eukaryotes dependent on DNA polymerase zeta and Rev1p. Philos Trans R Soc Lond B Biol Sci 356:41-6
Nelson, J R; Gibbs, P E; Nowicka, A M et al. (2000) Evidence for a second function for Saccharomyces cerevisiae Rev1p. Mol Microbiol 37:549-54
Gibbs, P E; Wang, X D; Li, Z et al. (2000) The function of the human homolog of Saccharomyces cerevisiae REV1 is required for mutagenesis induced by UV light. Proc Natl Acad Sci U S A 97:4186-91
Lawrence, C W; Gibbs, P E; Murante, R S et al. (2000) Roles of DNA polymerase zeta and Rev1 protein in eukaryotic mutagenesis and translesion replication. Cold Spring Harb Symp Quant Biol 65:61-9
Gibbs, P E; McGregor, W G; Maher, V M et al. (1998) A human homolog of the Saccharomyces cerevisiae REV3 gene, which encodes the catalytic subunit of DNA polymerase zeta. Proc Natl Acad Sci U S A 95:6876-80
Carty, M P; Lawrence, C W; Dixon, K (1996) Complete replication of plasmid DNA containing a single UV-induced lesion in human cell extracts. J Biol Chem 271:9637-47
Nelson, J R; Lawrence, C W; Hinkle, D C (1996) Thymine-thymine dimer bypass by yeast DNA polymerase zeta. Science 272:1646-9
Lawrence, C W; Hinkle, D C (1996) DNA polymerase zeta and the control of DNA damage induced mutagenesis in eukaryotes. Cancer Surv 28:21-31

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