The long term objective of the research described in this proposal is to understand the mechanisms by which various agents mutate living cells. The approach is to start with a mutagenic agent for which there is at east some information concerning the products induced in the DNA. This agent is used to mutagenize the cI gene of lambda phage, either as a prophage incorporated in the genome of the Escherichia coli host cell, as free phage in solution, or as isolated phage DNa in solution. The host cells are wild-type or have defined genetic defects affecting DNA replication and repair. The mutations produced are characterized by determining the induced changes in DNA base sequence. In some cases, the changes are inferred from reversion of mutants, from a collection of several hundred, of known sequence. In some cases, mutations induced in short synthetic polynucleotides; will be studied; for technical reasons these synthetic pieces will probably first be incorporated in the lacZ(alpha) gene in M13 phage. The results are used to determine mechanisms of mutagenesis. The agents to be studied in the period covered by this proposal are ultraviolet light, ionizing radiation, the base analog bromouracil, perhaps some agent which forms bulky adducts to DNA such as the aminopyrenes and, probably, some other agent which may come to our attention at a later time. Also, additional data will be collected, from this lab and from others now sequencing mutations in the lambda cI gene, on the utility of this system for studies in mutagenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028297-09
Application #
3275598
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1980-08-01
Project End
1990-07-31
Budget Start
1988-08-01
Budget End
1990-07-31
Support Year
9
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Sockett, H; Romac, S; Hutchinson, F (1991) DNA sequence changes in mutations induced by ultraviolet light in the gpt gene on the chromosome of Escherichia coli uvr+ and urvA cells. Mol Gen Genet 230:295-301
Sockett, H; Romac, S; Hutchinson, F (1991) Mutagenic specificity of N-methyl-N'-nitro-N-nitrosoguanidine in the gpt gene on a chromosome of Chinese hamster ovary cells and of Escherichia coli cells. Mol Gen Genet 227:252-9
Tindall, K R; Stein, J; Hutchinson, F (1988) Changes in DNA base sequence induced by gamma-ray mutagenesis of lambda phage and prophage. Genetics 118:551-60
Hutchinson, F; Yamamoto, K; Stein, J et al. (1988) Effect of photoreactivation on mutagenesis of lambda phage by ultraviolet light. J Mol Biol 202:593-601
Reed, J; Hutchinson, F (1987) Effect of the direction of DNA replication on mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine in adapted cells of Escherichia coli. Mol Gen Genet 208:446-9
Hutchinson, F (1987) A review of some topics concerning mutagenesis by ultraviolet light. Photochem Photobiol 45:897-903
Wood, R D; Hutchinson, F (1987) Ultraviolet light-induced mutagenesis in the Escherichia coli chromosome. Sequences of mutants in the cI gene of a lambda lysogen. J Mol Biol 193:637-41
Wood, R D; Stein, J (1986) Role of the RecF gene product in UV mutagenesis of lambda phage. Mol Gen Genet 204:82-4
Wood, R D; Hutchinson, F (1986) The role of pyrimidine dimers in mutagenesis by ultraviolet light. Prog Clin Biol Res 209A:191-8
Hutchinson, F; Wood, R D (1986) Mechanisms of mutagenesis of E. coli by ultraviolet light. Basic Life Sci 38:377-83

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