The goal of this application is to investigate the mechanism by which nitroaromatic carcinogens cause mutations. We shall focus on the major DNA adducts formed by the nitropyrenes, the primary mutagenic pollutants in diesel exhaust, but also plan to initiate work on the DNA adducts of 3-nitrobenzanthrone, a recently discovered potent mutagenic and carcinogenic contaminant in diesel. We hypothesize that a group of DNA adducts formed by these nitroarenes play important roles in the etiology of human cancer via the mutagenic effects in crucial sequences of cancer genes (e.g., p53). We also hypothesize that most of these mutations occur via error-prone bypass of the lesions by Y-family DNA polymerases. In order to test these hypotheses, we shall synthesize and characterize oligonucleotides containing the major guanine adducts of 1-nitropyene, 1,6- and 1,8-dinitropyere, and 3-nitrobenzanthrone in important gene sequences. These adduct-containing oligonucleotides will be used to construct single-stranded plasmid or viral vectors and mutagenicity and genotoxicity will be evaluated in Escherichia coli and mammalian cells. The effect of specific repair or replication proteins will be assessed by using repair-proficient and -deficient cell lines or inhibitors. The cellular studies will be complemented by investigating in vitro kinetics and fidelity of polymerization catalyzed by Dpo4 and four human Y- family DNA polymerases. Initially, we shall focus on the C8 guanine adduct of 1-nitropyrene. The translesion synthesis products will be sequenced to determine the mutagenic pattern caused by each DNA polymerase. Finally, duplex plasmid vectors with these adducts will be constructed, and progression of replication fork will be examined in human cell extracts to determine if each adduct is significant block to fork progression. Mutational types and frequencies in double-stranded DNA will be determined. Together, these studies will provide a deeper understanding of the mechanism of mutagenesis induced by the DNA adducts of these nitroaromatic carcinogens.

Public Health Relevance

Nitroaromatic compounds have been detected in a variety of environmental samples, including diesel exhaust. These compounds are potent mutagens and carcinogens, and it has been suggested that some of these pollutants play a role in human lung and breast cancer. Therefore, investigation of the mechanism by which the nitroaromatic carcinogens cause mutations has high significance to public health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES009127-14
Application #
8609028
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Shaughnessy, Daniel
Project Start
1998-02-01
Project End
2015-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
14
Fiscal Year
2014
Total Cost
$310,838
Indirect Cost
$63,070
Name
University of Connecticut
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
614209054
City
Storrs-Mansfield
State
CT
Country
United States
Zip Code
06269
Basu, Ashis K (2018) DNA Damage, Mutagenesis and Cancer. Int J Mol Sci 19:
Basu, Ashis K; Pande, Paritosh; Bose, Arindam (2017) Translesion Synthesis of 2'-Deoxyguanosine Lesions by Eukaryotic DNA Polymerases. Chem Res Toxicol 30:61-72
Tokarsky, E John; Wallenmeyer, Petra C; Phi, Kenneth K et al. (2017) Significant impact of divalent metal ions on the fidelity, sugar selectivity, and drug incorporation efficiency of human PrimPol. DNA Repair (Amst) 49:51-59
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Tokarsky, E John; Gadkari, Varun V; Zahurancik, Walter J et al. (2016) Pre-steady-state kinetic investigation of bypass of a bulky guanine lesion by human Y-family DNA polymerases. DNA Repair (Amst) 46:20-28
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Raper, Austin T; Gadkari, Varun V; Maxwell, Brian A et al. (2016) Single-Molecule Investigation of Response to Oxidative DNA Damage by a Y-Family DNA Polymerase. Biochemistry 55:2187-96
Bose, Arindam; Pande, Paritosh; Jasti, Vijay P et al. (2015) DNA polymerases ? and ? cooperatively perform mutagenic translesion synthesis of the C8-2'-deoxyguanosine adduct of the dietary mutagen IQ in human cells. Nucleic Acids Res 43:8340-51
Vyas, Rajan; Efthimiopoulos, Georgia; Tokarsky, E John et al. (2015) Mechanistic Basis for the Bypass of a Bulky DNA Adduct Catalyzed by a Y-Family DNA Polymerase. J Am Chem Soc 137:12131-42

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