CELLULAR DNA REPAIR MECHANISMS AND MUTAGENESIS

Project: Research project

Description

The objectives of the research are to understand mechanisms of DNA repair
and mutagenesis. The cellular process of repair is important in
maintenance of genetic information in correct context and expression
format. We now know that processes associated with repair of DNA actually
promote mutations in some cases. Mutagenesis of DNA can lead to genetic
disease and is a primary step in the pathway of carcinogenesis. We will investigate the regulation and function of the uvrC gene of E.
coli. This gene acts in excision repair of bulky damage in DNA. Our
approach will be to capitalize on a bank of recombinant DNA constructs
containing the structural gene with combinations of four 5' promoters. We
will analyze lexA regulation and do site-directed mutagenesis to determine
regulation. The overproduced gene product will be analyzed functionally in
an in vitro reaction to determine its role in excision. We will take advantage of a group of mutants of E. coli which can survive
without a functional DNA polymerase III. The conditional lethality of the
polC mutation is reversed by an extragenic mutation. These mutants offer a
unique resource to study the role of DNA polymerase III in repair and
mutagenesis. The mutants are more sensitive to hydrogen peroxide than
normal cells and we will use extracts from these mutants to develop an in
vitro repair assay for hydrogen peroxide.
StatusFinished
Effective start/end date1/1/7811/30/93

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $100,203.00
  • National Institutes of Health: $65,011.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $169,980.00

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Mutagenesis
DNA Repair
DNA Polymerase III
Genes
Escherichia coli
Recombinant DNA
Hydrogen Peroxide
Mutation
DNA
Carcinogenesis
Site-Directed Mutagenesis
Research
DNA Damage
Rec A Recombinases
DNA Polymerase I
Holoenzymes
Peroxides
Chromosomes
Cell Survival
Genetic Techniques

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)