Genetic recombination will be studied using yeast as an experimental system, taking advantage of both genetic and molecular approaches that are available in this organism, to gain insight into the various steps involved in the process of genetic recombination. Recombination assays will continue to be developed that focus on well-defined steps in the process. Mutations that block these steps will be searched for to eventually define the genes and the genetic interactions that function in recombination events. Specifically: (1) a known meiotic function in recombination events. Specifically: (1) a known meiotic hotspot near cyc3 will be analyzed to identify the DNA sequence(s) responsible and to determine the initiating lesion, (2) a plasmid will be designed to identify new meiotic hotspots and to serve as an assayable construct to select for trans-acting mutations that interact with known hotspots, (3) a recently developed mitotic heteroduplex assay from this lab will be used to assay the effect of known recombination and repair mutations as well as to select for new mutations that affect this process, (4) a plasmid that is sensitive to an in vivo double-strand- break will be designed for both selecting mutations that are essential for double-strand-break repair and for examining the intermediates of this process in wild type and mutant cells, (5) the effect of transcription and other local DNA perturbations on both meiotic and mitotic recombination will be determined and (6) an attempt will be made to identify yeast gene(s) involved in the resolution of Holliday structures by complementing a mutation in a bacteriophage T4 endonuclease that performs a similar function. It is hoped that understanding the steps of genetic recombination in yeast will help define the process in higher eukaryotic organisms as well.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034587-08
Application #
3285873
Study Section
Genetics Study Section (GEN)
Project Start
1984-07-01
Project End
1993-06-30
Budget Start
1991-07-01
Budget End
1993-06-30
Support Year
8
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Gangloff, S; Zou, H; Rothstein, R (1996) Gene conversion plays the major role in controlling the stability of large tandem repeats in yeast. EMBO J 15:1715-25
Smith, J; Rothstein, R (1995) A mutation in the gene encoding the Saccharomyces cerevisiae single-stranded DNA-binding protein Rfa1 stimulates a RAD52-independent pathway for direct-repeat recombination. Mol Cell Biol 15:1632-41
Rothstein, R; Gangloff, S (1995) Hyper-recombination and Bloom's syndrome: microbes again provide clues about cancer. Genome Res 5:421-6
Fasullo, M; Dave, P; Rothstein, R (1994) DNA-damaging agents stimulate the formation of directed reciprocal translocations in Saccharomyces cerevisiae. Mutat Res 314:121-33
McDonald, J P; Rothstein, R (1994) Unrepaired heteroduplex DNA in Saccharomyces cerevisiae is decreased in RAD1 RAD52-independent recombination. Genetics 137:393-405
Gangloff, S; McDonald, J P; Bendixen, C et al. (1994) The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase. Mol Cell Biol 14:8391-8
Bendixen, C; Sunjevaric, I; Bauchwitz, R et al. (1994) Identification of a mouse homologue of the Saccharomyces cerevisiae recombination and repair gene, RAD52. Genomics 23:300-3
Rothstein, R (1991) Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol 194:281-301
Bailis, A M; Rothstein, R (1990) A defect in mismatch repair in Saccharomyces cerevisiae stimulates ectopic recombination between homeologous genes by an excision repair dependent process. Genetics 126:535-47
Pachnis, V; Pevny, L; Rothstein, R et al. (1990) Transfer of a yeast artificial chromosome carrying human DNA from Saccharomyces cerevisiae into mammalian cells. Proc Natl Acad Sci U S A 87:5109-13

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