Abstract

The research proposed here is directed toward an understanding of the molecular mechanism of homologous genetic recombination. This goal is approached through the study of special sites in DNA promoting a high rate of recombination in their vicinity and through the identification and study of the activity of proteins interacting with these sites. The Chi recombinational hotspots of the bacterium Escherichia coil and the M26 recombinational hotspot of the fission yeast Schizosaccharomyces pombe will be studied in the proposed research. The interaction of Chi and RecBCD enzyme, which directly activates Chi, and the Mts1,2 protein, which binds to the M26 site, will be studied. The regional specificity of the S. pombe rec8, rec1O, and rec11 gene products, hypothesized to act at hotspots, will be studied. These studies will elucidate the molecular mechanisms by which recombinational hotspots act and thereby elucidate an important (rate-limiting) step in homologous recombination.
Specific aims are to: (A) determine the role of an essential RNA subunit of RecBCD enzyme in its interaction with Chi, (B) assess the mode of RecBCD-Chi interaction in E. coli cells, (C) determine the physical alteration of RecBCD upon cutting at Chi, (D) determine which strand of Chi is recognized by RecBCD, (B)test a model for Chi-stimulated conjugational and transductional recombination, (F)clone the mts1,2 genes and study mutants lacking the Mts1,2 protein, (G) determine the basis for the chromosomal context-dependence of M26, and (H) study the regional specificity of the Rec8, 10, and 11 activators of meiotic recombination. Recombination plays an important role in generating diversity among individuals in a population and, in some cases, among cells within an organism. Understanding the mechanism of recombination thus lends insight into the evolution and development of organisms. Aberrations of recombination may be responsible for certain chromosomal rearrangements associated with birth defects, hereditary diseases, and cancer.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM031693-16
Application #
2021942
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1982-07-01
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1997-12-31
Support Year
16
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109

  Publications

Fowler, Kyle R; Hyppa, Randy W; Cromie, Gareth A et al. (2018) Physical basis for long-distance communication along meiotic chromosomes. Proc Natl Acad Sci U S A 115:E9333-E9342
Amundsen, Susan K; Smith, Gerald R (2018) The RecB helicase-nuclease tether mediates Chi hotspot control of RecBCD enzyme. Nucleic Acids Res :
Nuckolls, Nicole L; Bravo Núñez, María Angélica; Eickbush, Michael T et al. (2017) wtf genes are prolific dual poison-antidote meiotic drivers. Elife 6:
Nambiar, Mridula; Smith, Gerald R (2016) Repression of harmful meiotic recombination in centromeric regions. Semin Cell Dev Biol 54:188-97
Amundsen, Susan K; Sharp, Jake W; Smith, Gerald R (2016) RecBCD Enzyme ""Chi Recognition"" Mutants Recognize Chi Recombination Hotspots in the Right DNA Context. Genetics 204:139-52
Taylor, Andrew F; Amundsen, Susan K; Smith, Gerald R (2016) Unexpected DNA context-dependence identifies a new determinant of Chi recombination hotspots. Nucleic Acids Res 44:8216-28
Fowler, Kyle R; Sasaki, Mariko; Milman, Neta et al. (2014) Evolutionarily diverse determinants of meiotic DNA break and recombination landscapes across the genome. Genome Res 24:1650-64
Zanders, Sarah E; Eickbush, Michael T; Yu, Jonathan S et al. (2014) Genome rearrangements and pervasive meiotic drive cause hybrid infertility in fission yeast. Elife 3:e02630
Taylor, Andrew F; Amundsen, Susan K; Guttman, Miklos et al. (2014) Control of RecBCD enzyme activity by DNA binding- and Chi hotspot-dependent conformational changes. J Mol Biol 426:3479-99
Hyppa, Randy W; Fowler, Kyle R; Cipak, Lubos et al. (2014) DNA intermediates of meiotic recombination in synchronous S. pombe at optimal temperature. Nucleic Acids Res 42:359-69

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