Eukaryotic cells rely on a conserved network of proteins to monitor the status of their genomes. This surveillance system detects double-strand breaks (DSBs) and transduces a checkpoint signal to the cell cycle machinery to coordinate cell division with DNA repair. Any defect in properly sensing and signaling aberrant DNA structures can lead to genome instability, which is a hallmark of cancer cells. This study will extend our efforts to use the fission yeast Schizosaccharomyces pombe to model mechanisms by which cells detect DSBs and activate a checkpoint response. The focus will be on two proteins: Crb2 and Ctp1. Crb2, an ortholog of the mammalian tumor suppressor/checkpoint protein 53BP1, belongs to a class of checkpoint proteins that also includes the tumor suppressor BRCA1, all of which have a C-terminal BRCT repeat. Like 53BP1 and BRCA1, Crb2 associates with DSBs, forming microscopically visible nuclear foci at sites of DNA damage. Ctp1, an ortholog of mammalian tumor suppressor/checkpoint protein CtIP, functions with the Mre11-Rad50-Nbs1 complex in resecting DSBs as the part of the initial step of DSB repair by homologous recombination. This study will: (1) examine the interactions between Crb2 and modified histones that define one mechanism of recruiting Crb2 to DSBs;(2) further characterize a mechanism responsible for histone modification- independent relocalization of Crb2 to DSBs that involves Cut5, the ortholog of human TopBP1;(3) define a mechanism in which fission yeast Tel1 (ortholog of human ATM tumor suppressor/checkpoint protein) activates Chk1 checkpoint kinase in the pre-resection phase of checkpoint signaling;and (4) determine how Ctp1 governs DSB repair during the cell cycle. Accomplishment of these aims will yield a more comprehensive mechanistic and functional understanding of the cellular response to DNA damage. A high-resolution picture of these mechanisms in fission yeast will aid in achieving a more detailed understanding of DNA damage responses and their connection to cancer in humans.

Public Health Relevance

DNA damage checkpoint proteins act as cellular firefighters: they rush to double-strand breaks in DNA where they help to repair the damage and send signals to stop cell division. This study will determine how one of these proteins, Crb2, is recruited to double-strand breaks. Insights gained from these studies will provide a more detailed understanding of DNA damage responses and their connection to cancer in humans.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA077325-14
Application #
8118764
Study Section
Molecular Genetics C Study Section (MGC)
Program Officer
Pelroy, Richard
Project Start
1998-04-20
Project End
2012-07-31
Budget Start
2011-08-01
Budget End
2012-07-31
Support Year
14
Fiscal Year
2011
Total Cost
$421,871
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Reubens, Michael C; Rozenzhak, Sophie; Russell, Paul (2017) Multi-BRCT Domain Protein Brc1 Links Rhp18/Rad18 and ?H2A To Maintain Genome Stability during S Phase. Mol Cell Biol 37:
Jensen, Kristi L; Russell, Paul (2016) Ctp1-dependent clipping and resection of DNA double-strand breaks by Mre11 endonuclease complex are not genetically separable. Nucleic Acids Res 44:8241-9
Guo, Lan; Ganguly, Abantika; Sun, Lingling et al. (2016) Global Fitness Profiling Identifies Arsenic and Cadmium Tolerance Mechanisms in Fission Yeast. G3 (Bethesda) 6:3317-3333
Petersen, Janni; Russell, Paul (2016) Growth and the Environment of Schizosaccharomyces pombe. Cold Spring Harb Protoc 2016:pdb.top079764
Sánchez, Arancha; Russell, Paul (2015) Ku stabilizes replication forks in the absence of Brc1. PLoS One 10:e0126598
Mejia-Ramirez, Eva; Limbo, Oliver; Langerak, Petra et al. (2015) Critical Function of ?H2A in S-Phase. PLoS Genet 11:e1005517
Sánchez, Arancha; Roguev, Assen; Krogan, Nevan J et al. (2015) Genetic Interaction Landscape Reveals Critical Requirements for Schizosaccharomyces pombe Brc1 in DNA Damage Response Mutants. G3 (Bethesda) 5:953-62
Wang, Lanfeng; Limbo, Oliver; Fei, Jia et al. (2014) Regulation of the Rhp26ERCC6/CSB chromatin remodeler by a novel conserved leucine latch motif. Proc Natl Acad Sci U S A 111:18566-71
Wei, Yi; Wang, Hai-Tao; Zhai, Yonggong et al. (2014) Mdb1, a fission yeast homolog of human MDC1, modulates DNA damage response and mitotic spindle function. PLoS One 9:e97028
Deshpande, Rajashree A; Williams, Gareth J; Limbo, Oliver et al. (2014) ATP-driven Rad50 conformations regulate DNA tethering, end resection, and ATM checkpoint signaling. EMBO J 33:482-500

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