Our long term goal is to understand the biology, biochemistry and genetics of the response of cells to DNA alkylation damage. Alkylating agents represent the most abundant class of chemical DNA damaging agents in our environment and they induce cell death and mutation. Since we are continuously exposed to these chemicals, and since certain alkylating agents are used for chemotherapy, it is important to understand exactly how cells respond these agents. It is becoming increasingly clear that organisms separated by enormous evolutionary distances employ similar proteins to protect against damage relentlessly inflicted upon their DNA, and we now know that bacteria, yeast and human cells induce the expression of specific sets of genes in response to DNA damage. Our studies on the response of E. coli, S. cerevisiae and human cells to DNA alkylation damage have become intertwined and are being executed in an integrated fashion. Much of the 'Proposed experiments are based on our findings that bacterial DNA repair functions can operate in eukaryotic cells, and vice versa, i.e., eukaryotic DNA repair functions can operate in bacterial cells. Specifically, we identified and isolated three eukaryotic genes by virtue of their ability to rescue E. coli form alkylation induced killing or nutation; a 3MeA DNA glycosylase, an 06-MeG DNA methyltransferase, and a gene of unknown function. These genes and their products will be characterized, paying particular attention to the specificities of the DNA repair activities and to the transcriptional regulation of the genes in response to DNA alkylation damage, and other types of DNA damage. In addition, a number of other eukaryotic genes (both yeast and human) whose products prevent alkylation induced killing and mutation will also be isolated and similarly characterized. It is anticipated that characterization of all these genes will not only deepen our understanding of the roles of known DNA repair enzymes in protecting cells against the effects of alkylating agents but will also uncover some hitherto unknown protective mechanisms. The health relatedness of this project lies in the fact that it will contribute to an understanding of some of the events that may lead to carcinogenesis (via somatic mutations and to the

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA055042-02
Application #
3199475
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1991-07-02
Project End
1995-04-30
Budget Start
1992-05-01
Budget End
1993-04-30
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Public Health
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Jordan, Jennifer J; Chhim, Sophea; Margulies, Carrie M et al. (2017) ALKBH7 drives a tissue and sex-specific necrotic cell death response following alkylation-induced damage. Cell Death Dis 8:e2947
Chaim, Isaac A; Nagel, Zachary D; Jordan, Jennifer J et al. (2017) In vivo measurements of interindividual differences in DNA glycosylases and APE1 activities. Proc Natl Acad Sci U S A 114:E10379-E10388
Calvo, Jennifer A; Allocca, Mariacarmela; Fake, Kimberly R et al. (2016) Parp1 protects against Aag-dependent alkylation-induced nephrotoxicity in a sex-dependent manner. Oncotarget 7:44950-44965
Fu, Dragony; Samson, Leona D; Hübscher, Ullrich et al. (2015) The interaction between ALKBH2 DNA repair enzyme and PCNA is direct, mediated by the hydrophobic pocket of PCNA and perturbed in naturally-occurring ALKBH2 variants. DNA Repair (Amst) 35:13-8
Meira, Lisiane B; Calvo, Jennifer A; Shah, Dharini et al. (2014) Repair of endogenous DNA base lesions modulate lifespan in mice. DNA Repair (Amst) 21:78-86
Ebrahimkhani, Mohammad R; Daneshmand, Ali; Mazumder, Aprotim et al. (2014) Aag-initiated base excision repair promotes ischemia reperfusion injury in liver, brain, and kidney. Proc Natl Acad Sci U S A 111:E4878-86
Mazumder, Aprotim; Tummler, Katja; Bathe, Mark et al. (2013) Single-cell analysis of ribonucleotide reductase transcriptional and translational response to DNA damage. Mol Cell Biol 33:635-42
Svensson, J Peter; Quirós Pesudo, Laia; McRee, Siobhan K et al. (2013) Genomic phenotyping by barcode sequencing broadly distinguishes between alkylating agents, oxidizing agents, and non-genotoxic agents, and reveals a role for aromatic amino acids in cellular recovery after quinone exposure. PLoS One 8:e73736
van Loon, Barbara; Samson, Leona D (2013) Alkyladenine DNA glycosylase (AAG) localizes to mitochondria and interacts with mitochondrial single-stranded binding protein (mtSSB). DNA Repair (Amst) 12:177-87
Fu, Dragony; Jordan, Jennifer J; Samson, Leona D (2013) Human ALKBH7 is required for alkylation and oxidation-induced programmed necrosis. Genes Dev 27:1089-100

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