Intracellular protein and DNA dynamics in competent Bacillus subtilis cells

Cell. 2005 Jul 15;122(1):73-84. doi: 10.1016/j.cell.2005.04.036.

Abstract

We have found that two DNA repair/recombination proteins localize differentially to the cell poles in competent Bacillus subtilis cells. RecA protein colocalized with competence protein ComGA, and its polar localization largely depended on ComGA and ComK activity, while RecN oscillated between the poles in a minute time frame, independent of any competence factor. Oscillation of RecN arrested upon addition of external DNA, suggesting that an interaction with incoming single-stranded (ss) DNA favors the localization of RecN at the pole containing the competence machinery. In agreement with this model, purified RecN protein showed ATP-dependent binding to ssDNA. Addition of DNA resulted in the formation of RecA threads emanating from the competence machinery. Our data show that in competent bacteria there exists a specifically positioned and dynamic ssDNA binding apparatus that accepts ssDNA taken up through the polar competence machinery and processes ssDNA for recombination with chromosomal DNA via extended RecA filaments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / cytology
  • Bacillus subtilis / growth & development
  • Bacillus subtilis / metabolism*
  • Bacterial Proteins / metabolism*
  • DNA Restriction Enzymes / metabolism
  • DNA, Bacterial / metabolism*
  • DNA, Single-Stranded / metabolism
  • Escherichia coli / cytology
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism
  • Green Fluorescent Proteins / metabolism
  • Microscopy, Fluorescence
  • Rec A Recombinases / metabolism
  • Transcription Factors / metabolism

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA, Single-Stranded
  • Transcription Factors
  • Green Fluorescent Proteins
  • comK protein, Bacillus subtilis
  • Rec A Recombinases
  • DNA Restriction Enzymes
  • RecN protein, Bacteria