The long-term goal of this proposal is to understand the biological roles of DNA helicase and helicase-related genes in recognizing and repairing spontaneous damage that occurs during the mitotic cell cycle. Mitotic DNA damage is dangerous to DNA replication, and can result in stalled or collapsed replication forks. The DNA damage is processed by lesion bypass pathways or homologous recombination, depending on the type of DNA damage. Homologous recombination (HR) must be tightly regulated to avoid genomic rearrangements. Defects in recombination, particularly from helicase and helicase-like genes, can result in blocked intermediates that trigger a cell cycle checkpoint. We will investigate the roles of helicase and helicase-like genes in repair and recombination, studying the control over with which pathway is used to repair damage during replication. The Srs2 DNA helicase and the SWI/SNF-like proteins Rad54, Rdh54 and Tid4 act against Rad51, an essential factor in homologous recombination. The Srs2 DNA helicase can regulate the type of repair pathway by disrupting a Rad51 filament, one of the first intermediates in HR. We will investigate how the balance between HR and postreplication repair (PRR) is achieved through the balance between the Srs2 helicase and Rad51 protein, and whether replication components have a role in this balance. Rad54, Rdh54 and Tid4 remove Rad51 from double stranded DNA results in a cell cycle checkpoint arrest and increased DNA rearrangements.
The specific aims are to understand why the nonessential Srs2 DNA helicase becomes essential in sister chromatid cohesion defective strains and why excess Rad51 protein is lethal, and the roles of the SWI/SNF-like proteins Rdh54 and Tid4 in maintenance of genomic stability. These goals will be achieved by genetic studies, examination of the consequences of Rad51 overexpression, microscopic examination of cells for markers of DNA damage, studies on the role of sister chromatid cohesion recombination repair. DNA repair mechanisms are essential for mutation avoidance and stability of the genetic information. When the repair mechanisms are defective, mutations and chromosome changes occur. These reveal normally silent mutations, which now drive cells to uncontrolled growth associated with human cancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM053738-12
Application #
7793446
Study Section
Molecular Genetics C Study Section (MGC)
Program Officer
Hagan, Ann A
Project Start
1997-05-01
Project End
2013-03-31
Budget Start
2010-04-01
Budget End
2013-03-31
Support Year
12
Fiscal Year
2010
Total Cost
$287,936
Indirect Cost
Name
New York University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Epshtein, Anastasiya; Potenski, Catherine J; Klein, Hannah L (2016) Increased Spontaneous Recombination in RNase H2-Deficient Cells Arises From Multiple Contiguous rNMPs and Not From Single rNMP Residues Incorporated by DNA Polymerase Epsilon. Microb Cell 3:248-254
Niu, Hengyao; Potenski, Catherine J; Epshtein, Anastasiya et al. (2016) Roles of DNA helicases and Exo1 in the avoidance of mutations induced by Top1-mediated cleavage at ribonucleotides in DNA. Cell Cycle 15:331-6
Potenski, Catherine J; Niu, Hengyao; Sung, Patrick et al. (2014) Avoidance of ribonucleotide-induced mutations by RNase H2 and Srs2-Exo1 mechanisms. Nature 511:251-4
Potenski, Catherine J; Klein, Hannah L (2014) How the misincorporation of ribonucleotides into genomic DNA can be both harmful and helpful to cells. Nucleic Acids Res 42:10226-34
Burgess, Rebecca C; Sebesta, Marek; Sisakova, Alexandra et al. (2013) The PCNA interaction protein box sequence in Rad54 is an integral part of its ATPase domain and is required for efficient DNA repair and recombination. PLoS One 8:e82630
Ferrari, Matteo; Nachimuthu, Benjamin Tamilselvan; Donnianni, Roberto Antonio et al. (2013) Tid1/Rdh54 translocase is phosphorylated through a Mec1- and Rad53-dependent manner in the presence of DSB lesions in budding yeast. DNA Repair (Amst) 12:347-55
Bermejo, Rodrigo; Capra, Thelma; Jossen, Rachel et al. (2011) The replication checkpoint protects fork stability by releasing transcribed genes from nuclear pores. Cell 146:233-46
Hoot, Samantha J; Zheng, Xiuzhong; Potenski, Catherine J et al. (2011) The role of Candida albicans homologous recombination factors Rad54 and Rdh54 in DNA damage sensitivity. BMC Microbiol 11:214
Chi, Peter; Kwon, YoungHo; Visnapuu, Mari-Liis et al. (2011) Analyses of the yeast Rad51 recombinase A265V mutant reveal different in vivo roles of Swi2-like factors. Nucleic Acids Res 39:6511-22
Saponaro, Marco; Callahan, Devon; Zheng, Xiuzhong et al. (2010) Cdk1 targets Srs2 to complete synthesis-dependent strand annealing and to promote recombinational repair. PLoS Genet 6:e1000858

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