This research investigates the relationship of DNA damage to processes involved in chemical mutagenesis. nucleoside adducts, derived from chemical carcinogens, chemotherapeutic agents and products of ionizing radiation, will be incorporated, site-specifically, into synthetic oligodeoxynucleotides. The mutagenic spectra of these lesions will be determined in mammalian cells and bacteria. New protecting groups will be developed that will permit site-specific incorporation of alkali-sensitive adducts into oligodeoxynucleotides. Synthetic methods for introducing crosslinks and pre-selected positions in duplex DNA will be established. Chemically-modified oligodeoxynucleotides will be used to explore molecular mechanisms by which DNA polymerases copy damaged templates and to delineate the process of DNA replication and repair, with special reference to chemical mutagenesis. Experimental systems will be developed which allow chemically-modified DNA to be integrated into chromosomal DNA and subsequently rescue for mutational analysis. Damaged DNA will be examined for its ability to enhance homologous recombination. The precise relationship between the number and location of DNA lesions and the induction of the SOS response will be determined. Physical methods, including 2D-NMR and molecular model building, will be used to establish the three dimensional structure of chemically-modified oligodeoxynucleotides in solution. The mutagenicity of the radiomimetic agent, bleomycin, could arise from the abasic sites, 3-phosphoglycolate residues, and/or nucleoside base propenals produced when this antibiotic reacts with DNA. We will examine the mutagenicity of the two structural lesions and characterize the adducts formed when nucleoside base propenals react with DNA. The significance of this research lies in establishing a molecular basis for the mutagenic effects of various types of DNA damage and in demonstrating its relationship to DNA repair. These phenomena are fundamental to an understanding of the primary events involved in chemical carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA017395-18
Application #
3164699
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1977-01-01
Project End
1993-12-31
Budget Start
1992-01-01
Budget End
1992-12-31
Support Year
18
Fiscal Year
1992
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Type
Schools of Medicine
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Li, Haoquan; Endutkin, Anton V; Bergonzo, Christina et al. (2017) DNA Deformation-Coupled Recognition of 8-Oxoguanine: Conformational Kinetic Gating in Human DNA Glycosylase. J Am Chem Soc 139:2682-2692
Li, Haoquan; Endutkin, Anton V; Bergonzo, Christina et al. (2016) A dynamic checkpoint in oxidative lesion discrimination by formamidopyrimidine-DNA glycosylase. Nucleic Acids Res 44:683-94
Gao, Shujuan; Honey, Sangeet; Futcher, Bruce et al. (2016) The non-homologous end-joining pathway of S. cerevisiae works effectively in G1-phase cells, and religates cognate ends correctly and non-randomly. DNA Repair (Amst) 42:1-10
Kuznetsov, Nikita A; Bergonzo, Christina; Campbell, Arthur J et al. (2015) Active destabilization of base pairs by a DNA glycosylase wedge initiates damage recognition. Nucleic Acids Res 43:272-81
Lukina, Maria V; Popov, Alexander V; Koval, Vladimir V et al. (2013) DNA damage processing by human 8-oxoguanine-DNA glycosylase mutants with the occluded active site. J Biol Chem 288:28936-47
Kuznetsov, Nikita A; Koval, Vladimir V; Zharkov, Dmitry O et al. (2012) Conformational dynamics of the interaction of Escherichia coli endonuclease VIII with DNA substrates. DNA Repair (Amst) 11:884-91
Bergonzo, Christina; Campbell, Arthur J; de los Santos, Carlos et al. (2011) Energetic preference of 8-oxoG eversion pathways in a DNA glycosylase. J Am Chem Soc 133:14504-6
Kirpota, Oleg O; Endutkin, Anton V; Ponomarenko, Michail P et al. (2011) Thermodynamic and kinetic basis for recognition and repair of 8-oxoguanine in DNA by human 8-oxoguanine-DNA glycosylase. Nucleic Acids Res 39:4836-50
Mechetin, Grigory V; Zharkov, Dmitry O (2011) Mechanism of translocation of uracil-DNA glycosylase from Escherichia coli between distributed lesions. Biochem Biophys Res Commun 414:425-30
Zharkov, Dmitry O; Mechetin, Grigory V; Nevinsky, Georgy A (2010) Uracil-DNA glycosylase: Structural, thermodynamic and kinetic aspects of lesion search and recognition. Mutat Res 685:11-20

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