Determination of the Paracoccus denitrificans SOS box.
By gel retardation experiments with crude cell extracts of Paracoccus denitrificans it was demonstrated that a protein specifically binds to the promoter of the P. denitrificans recA gene. PCR mutagenesis of the recA promoter showed that the GAACN7GAAC motif is required for the formation of the DNA-protein complex. This protein also binds to the GTTCN7GTTC motif, which is present in the promoter of the P. denitrificans uvrA gene. Mutational analysis of the promoter regions of both P. denitrificans recA and uvrA genes indicated that the GAACN7GAAC and GTTCN7GTTC sequences are required for DNA-damage-mediated induction of these two genes in vivo. Furthermore, the P. denitrificans recA gene was DNA-damage-inducible when introduced into cells of the phylogenetically related phototrophic bacterium Rhodobacter sphaeroides, although this inducibility was lost in mutants in the GAACN7GAAC motif. These results indicate that P. denitrificans possesses the same SOS box as R. sphaeroides, which, in agreement with previous work, is proposed as being the GTTCN7GTTC motif.
Del Rey
Alfonso
A
Diestra
Julio
J
de Henestrosa
Antonio R Fernández
ARF
Barbé
Jordi
J
D000251:Adenosine Triphosphatases / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009875:Operator Regions, Genetic / Q000235:genetics; D010231:Paracoccus denitrificans / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016364:Reading Frames; D011938:Rec A Recombinases / Q000235:genetics*; D012242:Rhodobacter sphaeroides / Q000235:genetics; D013014:SOS Response, Genetics / Q000235:genetics*; D013045:Species Specificity; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D011938:Rec A Recombinases; C068631:UvrA protein, E coli; D000251:Adenosine Triphosphatases