The long-term goal of the research proposed here is to determine the molecular mechanism of homologous genetic recombination. This objective is approached by a combination of genetic analysis of mutants altered in recombination and biochemical analysis of DNA from cells. This research uses the fission yeast Schizosaccharomyces pombe, a widely studied, highly tractable model organism with features similar to those of multicellular eukaryotes, including humans. The studies are focused on meiotic recombination, whose high rates of recombination facilitate both genetic and biochemical analyses.
Specific aims are to (1) determine how meiotic DNA double-strand break (DSB) formation is differentially regulated along chromosome arms by sister chromatid cohesins, linear element proteins, and protein kinases, (2) determine how meiotic DSB formation is repressed specifically in heterochromatic centromeres, and (3) determine how the Mus81-Eme1 Holliday junction resolvase is regulated in meiosis.
These aims will be attacked by a combination of genetic analysis of mutants, fluorescence microscopy of intracellular proteins, physical analysis of DNA intermediates from meiotic cells, and enzymology of isolated proteins. The results of these studies will elucidate the molecular mechanism of recombination as well as the controls on recombination that ensure that it occurs at the proper place along chromosomes to promote faithful segregation of homologs at the first meiotic division. Recombination is important for faithful meiotic chromosome segregation, for repair of DNA double-strand breaks, and for generating diversity at both the organismal and cellular levels. Aberrancies of recombination can generate chromosomal rearrangements, such as translocations, duplications, and deletions, which are often associated with or the cause of birth defects and cancers. Understanding the molecular mechanism of recombination is important in determining the causes of these diseases and possibly preventing them.

Public Health Relevance

Genetic recombination is important for the accurate repair of broken DNA, for formation of viable sex cells (eggs and sperm), and for enhancing diversity among cells and organisms. Failure of recombination can lead to birth defects and cancer, and understanding the molecular mechanism of recombination is important to prevent or possibly cure such diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM032194-31A1
Application #
8630559
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Janes, Daniel E
Project Start
1983-03-01
Project End
2017-11-30
Budget Start
2013-12-26
Budget End
2014-11-30
Support Year
31
Fiscal Year
2014
Total Cost
$419,074
Indirect Cost
$180,964
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
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
Cipak, Lubos; Polakova, Silvia; Hyppa, Randy W et al. (2014) Synchronized fission yeast meiosis using an ATP analog-sensitive Pat1 protein kinase. Nat Protoc 9:223-31
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
Fowler, Kyle R; Gutierrez-Velasco, Susana; Martin-Castellanos, Cristina et al. (2013) Protein determinants of meiotic DNA break hot spots. Mol Cell 49:983-96
Phadnis, Naina; Hyppa, Randy W; Smith, Gerald R (2011) New and old ways to control meiotic recombination. Trends Genet 27:411-21
Bonfils, Sandrine; Rozalen, Ana E; Smith, Gerald R et al. (2011) Functional interactions of Rec24, the fission yeast ortholog of mouse Mei4, with the meiotic recombination-initiation complex. J Cell Sci 124:1328-38
Reddy, Bharat D; Wang, Yu; Niu, Lifang et al. (2011) Elimination of a specific histone H3K14 acetyltransferase complex bypasses the RNAi pathway to regulate pericentric heterochromatin functions. Genes Dev 25:214-9
Ellermeier, Chad; Higuchi, Emily C; Phadnis, Naina et al. (2010) RNAi and heterochromatin repress centromeric meiotic recombination. Proc Natl Acad Sci U S A 107:8701-5
Milman, Neta; Higuchi, Emily; Smith, Gerald R (2009) Meiotic DNA double-strand break repair requires two nucleases, MRN and Ctp1, to produce a single size class of Rec12 (Spo11)-oligonucleotide complexes. Mol Cell Biol 29:5998-6005
Farah, Joseph A; Cromie, Gareth A; Smith, Gerald R (2009) Ctp1 and Exonuclease 1, alternative nucleases regulated by the MRN complex, are required for efficient meiotic recombination. Proc Natl Acad Sci U S A 106:9356-61

Showing the most recent 10 out of 64 publications