Abstract

The overall goal of this proposal is to study the genotoxicity and repair of oxidized DNA bases.
The specific aims are two. The first is to determine whether base excision repair (Dna glycosylase mediated) is selective in actively transcribed genes as has been shown for the nucleotide excision repair (multi-protein endo-nuclease) of UV dimers. We will study the repair of 5-Hydroxymethyluracil (HmUra) as an example of a lesion which does not block transcription. Secondly, we will study the phylogenetic distribution of the repair enzymes, HmUra-DNA glycosylase and 5-hydroxymethylcytosine (HmCyt)-DNA glycosylase. Both of these activities are present in mammalian tissue and are absent from bacteria. These are the only known DNA glycosylase activities showing this phylogenetic dissociation. A study of their phylogenetic distribution will clarify the susceptibility of methylcytosine and thymine residues to endogenous oxidative stress. Oxidative damage to DNA has been suggested to play a role in the etiology of cancer and aging. Our approach is a direct biochemical and phylogenetic study addressed at evaluating this hypothesis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA049869-02
Application #
3194185
Study Section
Metabolic Pathology Study Section (MEP)
Project Start
1989-04-15
Project End
1992-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
New York University
Department
Type
Schools of Medicine
DUNS #
004514360
City
New York
State
NY
Country
United States
Zip Code
10012

  Publications

Ocampo, Maria T A; Chaung, Wenren; Marenstein, Dina R et al. (2002) Targeted deletion of mNth1 reveals a novel DNA repair enzyme activity. Mol Cell Biol 22:6111-21
Boorstein, R J; Cummings Jr, A; Marenstein, D R et al. (2001) Definitive identification of mammalian 5-hydroxymethyluracil DNA N-glycosylase activity as SMUG1. J Biol Chem 276:41991-7
Marenstein, D R; Ocampo, M T; Chan, M K et al. (2001) Stimulation of human endonuclease III by Y box-binding protein 1 (DNA-binding protein B). Interaction between a base excision repair enzyme and a transcription factor. J Biol Chem 276:21242-9
Chheda, A D; Teebor, G W; Cunningham, R P (2000) Identification, characterization, and purification of DNA glycosylase/AP lyases by reductive crosslinking to 2'-deoxyribooligonucleotides containing specific base lesions. Methods 22:180-7
Hilbert, T P; Boorstein, R J; Kung, H C et al. (1996) Purification of a mammalian homologue of Escherichia coli endonuclease III: identification of a bovine pyrimidine hydrate-thymine glycol DNAse/AP lyase by irreversible cross linking to a thymine glycol-containing oligoxynucleotide. Biochemistry 35:2505-11
Zuo, S; Boorstein, R J; Cunningham, R P et al. (1995) Comparison of the effects of UV irradiation on 5-methyl-substituted and unsubstituted pyrimidines in alternating pyrimidine-purine sequences in DNA. Biochemistry 34:11582-90
Zuo, S; Boorstein, R J; Teebor, G W (1995) Oxidative damage to 5-methylcytosine in DNA. Nucleic Acids Res 23:3239-43
O'Donnell, R E; Boorstein, R J; Cunningham, R P et al. (1994) Effect of pH and temperature on the stability of UV-induced repairable pyrimidine hydrates in DNA. Biochemistry 33:9875-80
Lustig, M J; Cadet, J; Boorstein, R J et al. (1992) Synthesis of the diastereomers of thymidine glycol, determination of concentrations and rates of interconversion of their cis-trans epimers at equilibrium and demonstration of differential alkali lability within DNA. Nucleic Acids Res 20:4839-45
Levy, D D; Teebor, G W (1991) Site directed substitution of 5-hydroxymethyluracil for thymine in replicating phi X-174am3 DNA via synthesis of 5-hydroxymethyl-2'-deoxyuridine-5'-triphosphate. Nucleic Acids Res 19:3337-43

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