We propose to carry out a program of research on mutation in mammalian cells. As the target for mutations, these studies will use a small bacterial gene (gpt) introduced into mammalian cells as part of a retroviral shuttle vector and integrated into mammalian chromosomal DNA in a proviral form. We have recently developed this system and demonstrated its potential for providing information on mutations in mammalian cells at the level of specific base changes. The goal of the research is the elucidation of the molecular mechanisms of mutation in mammalian cells. The proposed studies will utilize shuttle vector transformed Gpt+ mouse cells and the large number of Gpt+ mutants that we have already isolated and characterized in terms of specific base changes, and other transformed lines and mutants to be isolated. The major specific aims are: (1) determine the base changes in mutations occuring spontaneously or as the result of mutagenesis by chemical or physical agents, (2) analyze the relationshp between nucleotide pool perturbation and mutagen-induced based changes in DNA, (3) determine the molecular mechanisms by which mutant genes in mammalian cells can revert; (4) isolate mammalian cell suppressor mutants, (5) determine if the mutational spectra for the gpt gene are different depending upon the recipient species or the site of integration of the shuttle vector, (6) examine mismatch repair in mammalian cells, (7) introduce the gpt gene in a shuttle vector into mice and analyze the molecular mechanisms of mutation in Gpt+ mice.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA031781-11
Application #
3169886
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1981-07-01
Project End
1993-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
11
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Type
Schools of Medicine
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Lall, L; Davidson, R L (1998) Sequence-directed base mispairing in human oncogenes. Mol Cell Biol 18:4659-69
Kresnak, M; Davidson, R L (1993) Dimethylsulfate methylation of guanine residues in mammalian DNA: inverse correlation between methylation susceptibility and mutagenesis by bromodeoxyuridine and thymidine. Somat Cell Mol Genet 19:589-98
Kresnak, M T; Davidson, R L (1992) Thymidine-induced mutations in mammalian cells: sequence specificity and implications for mutagenesis in vivo. Proc Natl Acad Sci U S A 89:2829-33
Rotstein, J B; Kresnak, M T; Samadashwily, G M et al. (1992) Analysis of sequence specificity of 5-bromodeoxyuridine-induced reversion in cells containing multiple copies of a mutant gpt gene. Somat Cell Mol Genet 18:179-88
Kresnak, M T; Davidson, R L (1991) Effects of flanking base sequences on 5-bromodeoxyuridine mutagenesis in mammalian cells. Somat Cell Mol Genet 17:399-410
Gelbert, L M; Wilson, M M; Davidson, R L (1990) Analysis of GPT activity in mammalian cells with a chromosomally integrated shuttle vector containing altered gpt genes. Somat Cell Mol Genet 16:173-84
Xu, F M; Greenspan, J A; Davidson, R L (1990) Replication-dependent mutagenesis by 5-bromodeoxyuridine: identification of base change and sequence effects on mutability. Somat Cell Mol Genet 16:477-86
Ashman, C R (1989) Retroviral shuttle vectors as a tool for the study of mutational specificity (base substitution/deletion/mutational hotspot). Mutat Res 220:143-9
Davidson, R L; Broeker, P; Ashman, C R (1988) DNA base sequence changes and sequence specificity of bromodeoxyuridine-induced mutations in mammalian cells. Proc Natl Acad Sci U S A 85:4406-10
Greenspan, J A; Xu, F M; Davidson, R L (1988) Molecular analysis of ethyl methanesulfonate-induced reversion of a chromosomally integrated mutant shuttle vector gene in mammalian cells. Mol Cell Biol 8:4185-9

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