Molecular genetic and biochemical analyses of a DNA repair gene from Serratia marcescens
Thesis/Dissertation
·
OSTI ID:5859137
In Escherichia coli, the SOS response and two 3-methyladenine DNA glycosylases (TagI and TagII) are required for repair of DNA damaged by alkylating agents such as methyl methanesulfonate (MMS). Mutations of the recA gene eliminate the SOS response. TagI and TagII are encoded by the tag and alkA genes, respectively. A gene (rpr) encoding 3-methyladenine DNA glycosylase activity was isolated from the Gram-negative bacterium Serratia marcescens. The gene, localized to a 1.5-kilobase pair SmaI-HindIII restriction fragment, was cloned into plasmid pUC18. The clone complemented E. coli tag alkA and recA mutations for MMS resistance. The rpr gene did not, however, complement recA mutations for resistance to ultraviolet light or the ability to perform homologous recombination reactions, nor did it complement E. coli ada or alkB mutations. Two proteins of molecular weights 42,000 and 16,000 were produced from the rpr locus. Analysis of deletion and insertion mutants of rpr suggested that the 42kD molecule is the active protein. The 16kD protein may either be a breakdown product of the 42kD species or may be encoded by another gene overlapping the reading frame of the rpr gene. Biochemical assays showed that the rpr gene product (Rpr) possesses 3-methyladenine DNA glycosylase activity.
- Research Organization:
- Louisiana State Univ. and Agricultural and Mechanical Coll., Baton Rouge, LA (USA)
- OSTI ID:
- 5859137
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550200* -- Biochemistry
550700 -- Microbiology
560130 -- Radiation Effects on Microorganisms
560300 -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
BACTERIA
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
DNA REPAIR
ELECTROMAGNETIC RADIATION
ENZYMES
ESCHERICHIA COLI
ESTERS
GENE MUTATIONS
GENES
GENETIC EFFECTS
GLYCOSYL HYDROLASES
HYDROLASES
METHYL METHANESULFONATE
MICROORGANISMS
MOLECULAR BIOLOGY
MUTAGENS
MUTATIONS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
RADIATIONS
RECOVERY
REPAIR
SERRATIA
SULFONIC ACID ESTERS
ULTRAVIOLET RADIATION
550700 -- Microbiology
560130 -- Radiation Effects on Microorganisms
560300 -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
BACTERIA
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
DNA REPAIR
ELECTROMAGNETIC RADIATION
ENZYMES
ESCHERICHIA COLI
ESTERS
GENE MUTATIONS
GENES
GENETIC EFFECTS
GLYCOSYL HYDROLASES
HYDROLASES
METHYL METHANESULFONATE
MICROORGANISMS
MOLECULAR BIOLOGY
MUTAGENS
MUTATIONS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
RADIATIONS
RECOVERY
REPAIR
SERRATIA
SULFONIC ACID ESTERS
ULTRAVIOLET RADIATION