Abstract

The objective of this research is to continue the development and application of a model system for investigating the genetic effects of specific forms of chemical damage to the genome. In the work done over the past five years, we developed a strategy for building 06-methylguanine, as representative of a model carcinogen-DNA adduct, into a unique site in the genome of a bacterial virus. This adduct was replicated in vivo, and the qualitative and quantitative features of its mutagenesis were defined, as well as factors that influenced its repair. We have preliminary evidence that the strategy we have developed is of more general value for building into DNA other carcinogen-DNA adducts, even adducts that are bulky and/or chemically labile. In this application we are proposing to investigate the genetic effects of several of the DNA lesions formed by carcinogenic alkylating agents and by the potent liver carcinogen, aflatoxin B1.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA043066-03
Application #
3184955
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1987-07-01
Project End
1990-05-31
Budget Start
1989-06-01
Budget End
1990-05-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Massachusetts Institute of Technology
Department
Type
Other Specialized Schools
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139

  Publications

Dosanjh, M K; Essigmann, J M; Goodman, M F et al. (1990) Comparative efficiency of forming m4T.G versus m4T.A base pairs at a unique site by use of Escherichia coli DNA polymerase I (Klenow fragment) and Drosophila melanogaster polymerase alpha-primase complex. Biochemistry 29:4698-703
Ellison, K S; Dogliotti, E; Connors, T D et al. (1989) Site-specific mutagenesis by O6-alkylguanines located in the chromosomes of mammalian cells: influence of the mammalian O6-alkylguanine-DNA alkyltransferase. Proc Natl Acad Sci U S A 86:8620-4
Essigmann, J M; Basu, A K; Loechler, E L (1989) Mutagenic specificity of alkylated and oxidized DNA bases as determined by site-specific mutagenesis. Ann Ist Super Sanita 25:155-61
Singer, B; Chavez, F; Goodman, M F et al. (1989) Effect of 3' flanking neighbors on kinetics of pairing of dCTP or dTTP opposite O6-methylguanine in a defined primed oligonucleotide when Escherichia coli DNA polymerase I is used. Proc Natl Acad Sci U S A 86:8271-4
Couto, L B; Chaudhuri, I; Donahue, B A et al. (1989) Separation of the SOS-dependent and SOS-independent components of alkylating-agent mutagenesis. J Bacteriol 171:4170-7
Ellison, K S; Dogliotti, E; Essigmann, J M (1989) Construction of a shuttle vector containing a single O6-methylguanine: a probe for mutagenesis in mammalian cells. Mutat Res 220:93-100
Wilson, V L; Basu, A K; Essigmann, J M et al. (1988) O6-alkyldeoxyguanosine detection by 32P-postlabeling and nucleotide chromatographic analysis. Cancer Res 48:2156-61