Involvement of Escherichia coli DNA polymerase II in response to oxidative damage and adaptive mutation

Escarceller, M., Hicks, J., Gudmundsson, G., Trump, G., Touati, D., Lovett, S., Foster, P. L., McEntee, K., Goodman, M. F. (1994) Involvement of Escherichia coli DNA polymerase II in response to oxidative damage and adaptive mutation. Journal of Bacteriology, 176 (20). pp. 6221-6228. ISSN 00219193 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/7928992

Abstract

DNA polymerase II (Pol II) is regulated as part of the SOS response to DNA damage in Escherichia coli. We examined the participation of Pol II in the response to oxidative damage, adaptive mutation, and recombination. Cells lacking Pol II activity (polBΔ1 mutants) exhibited 5- to 10-fold-greater sensitivity to mode 1 killing by H 2O 2 compared with isogenic polB + cells. Survival decreased by about 15-fold when polB mutants containing defective superoxide dismutase genes, sodA and sodB, were compared with polB + sodA sodB mutants. Resistance to peroxide killing was restored following P1 transduction of polB cells to polB + or by conjugation of polB cells with an F' plasmid carrying a copy of polB +. The rate at which Lac + mutations arose in Lac - cells subjected to selection for lactose utilization, a phenomenon known as adaptive mutation, was increased threefold in polB backgrounds and returned to wild-type rates when polB cells were transduced to polB +. Following multiple passages of polB cells or prolonged starvation, a progressive loss of sensitivity to killing by peroxide was observed, suggesting that second-site suppressor mutations may be occurring with relatively high frequencies. The presence of suppressor mutations may account for the apparent lack of a mutant phenotype in earlier studies. A well- established polB strain, a dinA Mu d(Ap(r) lac) fusion (GW1010), exhibited wild-type (Pol II +) sensitivity to killing by peroxide, consistent with the accumulation of second-site suppressor mutations. A high-titer anti-Pol II polyclonal antibody was used to screen for the presence of Pol II in other bacteria and in the yeast Saccharomyces cerevisiae. Cross-reacting material was found in all gram-negative strains tested but was not detected in gram- positive strains or in S. cerevisiae. Induction of Pol II by nalidixic acid was observed in E. coli K-12, B, and C, in Shigella flexneri, and in Salmonella typhimurium.

Item Type: Paper
Uncontrolled Keywords: dna directed dna polymerase alpha hydrogen peroxide lactose nalidixic acid polyclonal antibody superoxide dismutase article bacterial mutation bactericidal activity bacterium transduction dna damage dna recombination dna repair enzyme induction escherichia coli nonhuman oxidative stress priority journal salmonella typhimurium shigella flexneri suppressor gene Adaptation, Physiological DNA Polymerase II Mutagenesis Oxidation-Reduction Phenotype Recombination, Genetic Sequence Homology SOS Response (Genetics) Support, U.S. Gov't, P.H.S. Variation (Genetics)
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > DNA polymerase
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
organism description > bacteria
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
organism description > bacteria > escherichia coli
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
CSHL Authors:
Communities: CSHL labs > Hicks lab
Depositing User: Matt Covey
Date: 1994
Date Deposited: 09 Aug 2013 20:55
Last Modified: 12 Aug 2013 14:51
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28501

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving