The repair of lesions in DNA is accomplished by a variety of mechanisms in mammalian cells. Perhaps the two most well-studied of these pathways are nucleotide excision repair and base excision repair. In contrast, recombinational repair pathways that repair double-strand breaks and interstrand crosslinks have not, as yet, been as thoroughly investigated. For interstrand crosslink repair in particular, a major impediment to progress has been the lack of an in vitro cell-free assay which would allow a biochemical analysis of this pathway. In this proposal, we show that such an assay has been developed with mammalian cell-free extracts. Results from the assay correlate very strongly with the known sensitivity of various cell lines to crosslinking agents. This assay will be utilized to determine the factors required in the assay, to investigate the mechanisms of recombinational repair of interstrand crosslinks, and to begin the biochemical fractionation of factors that are involved in this pathway. Finally, in addition to these studies, two novel human genes, that are homologues of the yeast RAD51/RAD55/RAD57 family, have been obtained from a private sequence database. These new RecA-like genes will be characterized with particular emphasis on their possible role in crosslink repair.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA075160-02
Application #
2896102
Study Section
Radiation Study Section (RAD)
Program Officer
Pelroy, Richard
Project Start
1998-08-01
Project End
2001-07-31
Budget Start
1999-08-01
Budget End
2000-07-31
Support Year
2
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Texas MD Anderson Cancer Center
Department
Genetics
Type
Other Domestic Higher Education
DUNS #
001910777
City
Houston
State
TX
Country
United States
Zip Code
77030
Zhang, Nianxiang; Liu, Xiuping; Li, Lei et al. (2007) Double-strand breaks induce homologous recombinational repair of interstrand cross-links via cooperation of MSH2, ERCC1-XPF, REV3, and the Fanconi anemia pathway. DNA Repair (Amst) 6:1670-8
Lu, Xiaoyan; Legerski, Randy J (2007) The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage. Biochem Biophys Res Commun 354:968-74
Zheng, Huyong; Wang, Xin; Legerski, Randy J et al. (2006) Repair of DNA interstrand cross-links: interactions between homology-dependent and homology-independent pathways. DNA Repair (Amst) 5:566-74
Zhang, Nianxiang; Kaur, Ramandeep; Lu, Xiaoyan et al. (2005) The Pso4 mRNA splicing and DNA repair complex interacts with WRN for processing of DNA interstrand cross-links. J Biol Chem 280:40559-67
Lu, Xiaoyan; Zhang, Nianxiang; Vasquez, Karen et al. (2005) Repair of psoralen interstrand cross-links in Xenopus laevis egg extracts is highly mutagenic. Biochem Biophys Res Commun 336:69-75
Zhang, Nianxiang; Lu, Xiaoyan; Legerski, Randy J (2003) Partial reconstitution of human interstrand cross-link repair in vitro: characterization of the roles of RPA and PCNA. Biochem Biophys Res Commun 309:71-8
Zheng, Huyong; Wang, Xin; Warren, Amy J et al. (2003) Nucleotide excision repair- and polymerase eta-mediated error-prone removal of mitomycin C interstrand cross-links. Mol Cell Biol 23:754-61
Zhang, Xiaoshan; Richie, Christopher; Legerski, Randy J (2002) Translation of hSNM1 is mediated by an internal ribosome entry site that upregulates expression during mitosis. DNA Repair (Amst) 1:379-90
Zhang, Nianxiang; Lu, Xiaoyan; Zhang, Xiaoshan et al. (2002) hMutSbeta is required for the recognition and uncoupling of psoralen interstrand cross-links in vitro. Mol Cell Biol 22:2388-97
Richie, Christopher T; Peterson, Carolyn; Lu, Tao et al. (2002) hSnm1 colocalizes and physically associates with 53BP1 before and after DNA damage. Mol Cell Biol 22:8635-47

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