The long term goal of this proposal is to understand the biology, biochemistry and genetics of cellular responses to DNA alkylation damage. Alkylating agents represent the most abundant class of chemical DNA damaging agents in our environment and they are even present as natural cellular metabolites. These agents are capable of inducing cell death, mutation, chromosome damage, and cancer. Since we are continuously exposed to these chemicals, and since certain alkylating agents are used for the chemotherapeutic treatment of cancer patients, it is important to understand exactly how cells respond to these agents. The mechanisms that cells employ to defend against DNA damaging agents have been highly conserved, and we now know that bacteria, yeast and mammalian cells employ similar strategies to prevent the induction of cell death and mutation. In recent years Dr. Samson has identified a number of eukaryotic DNA alkylation repair genes encoding DNA glycosylases and DNA methyltransferases. This proposal presents an integrated plan, designed to enhance our understanding of how cells respond to alkylating agents. The plan was designed to encompass research at various levels of development; i.e., at the level of identifying new pathways that protect cells against alkylating agents, at the level of further characterizing recently discovered pathways, and at the level of detailed characterization of pathways that have been under study for several years. This purposefully broad-based plan results in a large, diverse proposal which should enhance our understanding of the eukaryotic response to DNA damage.
The Specific Aims i nclude the following: a detailed genetic and biochemical analysis of the transcriptional regulation of the S. cerevisiae MAG 3MeA DNA glycosylase gene and several other DNA repair and metabolism genes; further analysis of the in vivo role MAG; the generation and characterization of cell lines and whole animals (mice) with altered levels of DNA alkylation repair enzymes; the isolation of new eukaryotic genes involved in the response of cells to DNA alkylation damage. The health relatedness of this project lies in the fact that it will contribute to an understanding of some of the events that lead to carcinogenesis and to the appearance of mutations in germ line cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA055042-07
Application #
2376861
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1991-07-02
Project End
2000-02-29
Budget Start
1997-03-01
Budget End
1998-02-28
Support Year
7
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Harvard University
Department
Other Basic Sciences
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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