Intracellular protein and DNA dynamics in competent Bacillus subtilis cells.
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.
Kidane
Dawit
D
Institut für Mikrobiologie, Fachbereich Biologie II, Universität Freiburg, Verfügungsgebäude, Stefan-Meier-Str. 19, 79104 Freiburg, Germany.
Graumann
Peter L
PL
D001412:Bacillus subtilis / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011938:Rec A Recombinases / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; C087449:comK protein, Bacillus subtilis; D011938:Rec A Recombinases; D004262:DNA Restriction Enzymes; C047879:RecN protein, Bacteria