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The SbcCD complex ofDeinococcus radioduranscontributes to radioresistance and DNA strand break repairin vivoand exhibits Mre11–Rad50 type activity in vitro

Authors
Journal
DNA Repair
1568-7864
Publisher
Elsevier
Publication Date
Volume
9
Issue
5
Identifiers
DOI: 10.1016/j.dnarep.2010.01.012
Keywords
  • Deinococcus
  • Dsb Repair
  • Homologous Recombination
  • Mre11–Rad50 Activity
  • Radioresistance
  • Sbccd Complex
Disciplines
  • Biology

Abstract

Abstract Deinococcus radiodurans lacks a homologue of the recB and recC genes, and the sbcA/B genes, of Escherichia coli. Thus, DNA strand break repair in Deinococcus proceeds by pathways that do not utilize these proteins. Unlike E. coli, the absence of recBC and sbcA/sbcB, and presence of only sbcC and sbcD in Deinococcus, indicates an enigmatic role of SbcCD in this bacterium. Studies on sbcCD mutation in Deinococcus showed nearly a 100-fold increase in gamma radiation sensitivity as compared to wild type. The mutant showed a higher rate of in vivo DNA degradation during the post-irradiation recovery period that corresponds to the RecA-dependent DSB repair phase. These cells showed a typical NotI pattern of DNA reassembly during the early phase of DSB repair, but were defective for the subsequent RecA-dependent phase II of DSB repair. Hydrogen peroxide had no effect on cell survival of the mutant. While its tolerance to higher doses of UVC and mitomycin C was significantly decreased as compared to wild type. Purified recombinant SbcCD proteins showed single-stranded endonuclease and 3′ → 5′ double-stranded DNA exonuclease activities similar to that of the Mre11–Rad50 complex, which is required for DNA strand break repair in higher organisms. These results suggested that the Mre11–Rad50 type nuclease activity of SbcCD proteins contributes to the radiation resistance of D. radiodurans perhaps by promoting the RecA-dependent DSB repair required for polyploid genome maturation.

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