DNA interstrand crosslinks (ICLs) present a formidable block to DNA metabolic processes and must be repaired for cell survival. While much work has been done to define the mechanisms of ICL repair in bacteria and yeast, their processing in mammalian cells is not clearly defined. In the previous funding period, we demonstrated an nucleotide excision repair (NER)-dependent, error-prone repair of triplex-directed psoralen ICLs in mammalian cells. We also found that mismatch repair (MMR) proteins are involved in the response to and repair of psoralen ICLs in an error-free process. Our working hypothesis in this renewal application is that NER and mismatch repair (MMR) proteins interact in the recognition and initial processing of ICLs, while MMR proteins, independently of NER factors, are involved in ICL-induced cell-cycle regulation and apoptosis. The long-term objectives of the proposed research are to elucidate molecular mechanisms involved in the removal of DNA ICLs from the mammalian genome, to identify interactions among proteins from HR, MMR, and NER pathways in doing so, and to determine the roles of these proteins in other cellular responses to these lesions. Specifically we propose to: 1) test the hypothesis that proteins from the NER and MMR pathways interact during recognition and initial processing of ICLs;2) test the proposal that MLH1 functions in cellular checkpoint and apoptosis responses to DNA ICLs;3) target DNA ICLs to specific genomic sites in mutant mammalian cell lines to determine roles for DNA repair and recombination gene products in processing ICLs;and 4) assess the potential of targeted ICLs as antiproliferative therapeutic agents. The results obtained from these studies will provide valuable information to develop improved targeted strategies to control human cancers using ICL-inducing agents.

Public Health Relevance

This Research Project is part of a multicomponent Program Project with the theme of understanding the processing of complex DNA damage by mammalian cells. The significance to human health is to generate new knowledge and paradigms for modeling DNA repair of DNA interstrand crosslinks (ICLs), to improve therapy using ICL-inducing compounds, and to identify new therapeutic targets for cancer treatment.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
2P01CA097175-06A1
Application #
7781951
Study Section
Special Emphasis Panel (ZCA1-GRB-S (O1))
Project Start
2010-02-01
Project End
2014-12-31
Budget Start
2010-02-01
Budget End
2010-12-31
Support Year
6
Fiscal Year
2010
Total Cost
$203,998
Indirect Cost
Name
University of Texas MD Anderson Cancer Center
Department
Type
DUNS #
800772139
City
Houston
State
TX
Country
United States
Zip Code
77030
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Lange, Sabine S; Tomida, Junya; Boulware, Karen S et al. (2016) The Polymerase Activity of Mammalian DNA Pol ? Is Specifically Required for Cell and Embryonic Viability. PLoS Genet 12:e1005759
Mukherjee, Anirban; Vasquez, Karen M (2016) HMGB1 interacts with XPA to facilitate the processing of DNA interstrand crosslinks in human cells. Nucleic Acids Res 44:1151-60
Wood, Richard D; DoubliƩ, Sylvie (2016) DNA polymerase ? (POLQ), double-strand break repair, and cancer. DNA Repair (Amst) 44:22-32
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Manandhar, Mandira; Boulware, Karen S; Wood, Richard D (2015) The ERCC1 and ERCC4 (XPF) genes and gene products. Gene 569:153-61
Zahn, Karl E; Averill, April M; Aller, Pierre et al. (2015) Human DNA polymerase ? grasps the primer terminus to mediate DNA repair. Nat Struct Mol Biol 22:304-11

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