Recombinational repair protects against the lethal effects of DNA damaging agents. However, recombination can also provide deletions and gene conversion events that contribute to tumorigenesis. Alkylating agents damage DNA and are toxic and carcinogenic. These agents are abundant in our environment and produced in our cells as normal cellular metabolites. Ironically, these agents are also used to treat cancer. 3- Methyladenine (3MeA) DNA glycosylases protect cells from alkylation induced killing by releasing a broad spectrum of damaged bases from the DNA. The goals of this study are to determine how DNA repair influences cellular susceptibility to potentially deleterious recombination events, and to dissect the molecular mechanism of damage induced recombination. Our hypothesis is that DNA normally suppresses recombination by removing recombinogenic lesions, but that repair enzymes can also stimulate recombination if they convert damaged base into a more recombinogenic intermediate.
Specific Aim I is to explore the molecular basis for the effects of 3MeA DNA glycosylase activity on homologous recombination in yeast.
Specific Aim II is to develop a fluorescence detection system for quantifying the proportions of particular recombination events. This tool will help to reveal how alkyl lesions promote recombination in mammals, and whether or not alkylation damage is likely to underlie spontaneous recombination events in people.
Specific Aim III is to apply this detection system to isogenic mammalian cells that carry specific DNA repair defects in order to reveal the role of these pathways in prevent recombination events. The ability of 3 MeA DNA glycosylase activity to protect against homologous recombination in mice will be determined. The mechanisms by which specific alkyl lesions and complex cancer chemotherapeutics stimulate recombination will be explored. The broad long-term objective of this work is to reveal the molecular mechanisms of alkylation damage induced recombination in order to improve our ability to understand the causes of mitotic recombination in people, to identify individuals who have increased susceptibility to cancer, and to improve our ability to develop better anti-cancer therapies.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA079827-01
Application #
2733431
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Okano, Paul
Project Start
1999-02-25
Project End
2003-01-31
Budget Start
1999-02-25
Budget End
2000-01-31
Support Year
1
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Miscellaneous
Type
Other Domestic Higher Education
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Klapacz, Joanna; Pottenger, Lynn H; Engelward, Bevin P et al. (2016) Contributions of DNA repair and damage response pathways to the non-linear genotoxic responses of alkylating agents. Mutat Res Rev Mutat Res 767:77-91
Kiraly, Orsolya; Gong, Guanyu; Olipitz, Werner et al. (2015) Inflammation-induced cell proliferation potentiates DNA damage-induced mutations in vivo. PLoS Genet 11:e1004901
Kiraly, Orsolya; Gong, Guanyu; Roytman, Megan D et al. (2014) DNA glycosylase activity and cell proliferation are key factors in modulating homologous recombination in vivo. Carcinogenesis 35:2495-502
Wiktor-Brown, Dominika M; Sukup-Jackson, Michelle R; Fakhraldeen, Saja A et al. (2011) p53 null fluorescent yellow direct repeat (FYDR) mice have normal levels of homologous recombination. DNA Repair (Amst) 10:1294-9
Mutamba, James T; Svilar, David; Prasongtanakij, Somsak et al. (2011) XRCC1 and base excision repair balance in response to nitric oxide. DNA Repair (Amst) 10:1282-93
Olipitz, Werner; Hembrador, Sheena; Davidson, Matthew et al. (2010) Development and characterization of a novel variable low dose-rate irradiator for in vivo mouse studies. Health Phys 98:727-34
Kim, Ki Hean; Ragan, Timothy; Previte, Michael J R et al. (2007) Three-dimensional tissue cytometer based on high-speed multiphoton microscopy. Cytometry A 71:991-1002
Koturbash, I; Rugo, R E; Hendricks, C A et al. (2006) Irradiation induces DNA damage and modulates epigenetic effectors in distant bystander tissue in vivo. Oncogene 25:4267-75
Nowosielska, Anetta; Smith, Stephen A; Engelward, Bevin P et al. (2006) Homologous recombination prevents methylation-induced toxicity in Escherichia coli. Nucleic Acids Res 34:2258-68
Wiktor-Brown, Dominika M; Hendricks, Carrie A; Olipitz, Werner et al. (2006) Age-dependent accumulation of recombinant cells in the mouse pancreas revealed by in situ fluorescence imaging. Proc Natl Acad Sci U S A 103:11862-7

Showing the most recent 10 out of 19 publications