Homologous recombination is required during meiosis to promote accurate segregation of homologous chromosomes at the first meiotic division. Several proteins have been identified that re required for MEIOTIC RECOMBINATION in the YEAST Saccharomyces, but the specific role of these proteins in recombination is largely unknown. The long term goals of this work are to determine the molecular mechanism of meiotic recombination and how the cell cycle machinery coordinates recombination with other meiotic events. This project focuses on one meiotic protein, Dmcl, that is required for meiotic recombination. Dmc1 and its relative Rad51 share structural and functional homology to RecA protein. RecA is the central protein required fo recombinogenic REPAIR OF DNA DAMAGE in E. Coli. Yeast Rad51, and probably Dmc1 as well, share RecAs ability to promote strand exchange between two molecules. Dmcl protein will be purified and characterized to determine what activities it shares with RecA. The formation of recombination products requires numerous enzymatic activities in addition to those displayed by RecA. Many of the proteins required for meiotic recombination form a multi protein complex. Affinity chromatography and tandem mass spectroscopy will be used to identify proteins that form complexes with Dmcl during recombination. Time-course and mutational analyses will be used to characterize overall progress in the assembly and disassembly of functional recombination complexes. In addition, confocal microscopy will be used to examine individual recombination events in living cells using green fluorescent protein. The functions of Rad51 and Dmc1 are only partially redundant. We propose mutant screens designed to separate the redundant function of Dmc1 from its unique functions. These mutations will then be used to characterize the mechanistic features of meiotic recombination that distinguish it from mitotic recombinational repair. The function of Dmc1 is monitored by a checkpoint control mechanism that ensures recombination is complete before chromosome segregation occurs. We recently showed the genes required for checkpoint control have additional functions in synapsis and suppression of ectopic recombination. The relationship between these newly identified functions and checkpoint control will be examined.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM050936-09
Application #
6519580
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Program Officer
Anderson, Richard A
Project Start
1994-04-01
Project End
2003-03-31
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
9
Fiscal Year
2002
Total Cost
$298,478
Indirect Cost
Name
University of Chicago
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
Chan, Yuen-Ling; Zhang, Annie; Weissman, Benjamin P et al. (2018) RPA resolves conflicting activities of accessory proteins during reconstitution of Dmc1-mediated meiotic recombination. Nucleic Acids Res :
Gataulin, Daniil V; Carey, Jeffrey N; Li, Junya et al. (2018) The ATPase activity of E. coli RecA prevents accumulation of toxic complexes formed by erroneous binding to undamaged double stranded DNA. Nucleic Acids Res 46:9510-9523
Chan, Yuen-Ling; Bishop, Douglas K (2018) Purification of Saccharomyces cerevisiae Homologous Recombination Proteins Dmc1 and Rdh54/Tid1 and a Fluorescent D-Loop Assay. Methods Enzymol 600:307-320
Grubb, Jennifer; Brown, M Scott; Bishop, Douglas K (2015) Surface Spreading and Immunostaining of Yeast Chromosomes. J Vis Exp :e53081
Mason, Jennifer M; Dusad, Kritika; Wright, William Douglass et al. (2015) RAD54 family translocases counter genotoxic effects of RAD51 in human tumor cells. Nucleic Acids Res 43:3180-96
Shinohara, Miki; Hayashihara, Kayoko; Grubb, Jennifer T et al. (2015) DNA damage response clamp 9-1-1 promotes assembly of ZMM proteins for formation of crossovers and synaptonemal complex. J Cell Sci 128:1494-506
Brown, M Scott; Grubb, Jennifer; Zhang, Annie et al. (2015) Small Rad51 and Dmc1 Complexes Often Co-occupy Both Ends of a Meiotic DNA Double Strand Break. PLoS Genet 11:e1005653
Joshi, Neeraj; Brown, M Scott; Bishop, Douglas K et al. (2015) Gradual implementation of the meiotic recombination program via checkpoint pathways controlled by global DSB levels. Mol Cell 57:797-811
Chan, Yuen-Ling; Brown, M Scott; Qin, Daoming et al. (2014) The third exon of the budding yeast meiotic recombination gene HOP2 is required for calcium-dependent and recombinase Dmc1-specific stimulation of homologous strand assimilation. J Biol Chem 289:18076-86
Lao, Jessica P; Cloud, Veronica; Huang, Chu-Chun et al. (2013) Meiotic crossover control by concerted action of Rad51-Dmc1 in homolog template bias and robust homeostatic regulation. PLoS Genet 9:e1003978

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