Functional analysis of the recA promoter of Rhodobacter capsulatus.
Expression of the Rhodobacter capsulatus recA gene is inducible by DNA damage. By using primer extension and serial deletion, we have identified the promoter of the R. capsulatus recA gene. Electrophoretic mobility-shift assays experiments have shown that a protein binds to a region of the R. capsulatus recA promoter containing the imperfect palindromic TTGTACTCATACCATGAGAACAA, which is centered on position-8 with respect to the transcriptional starting site. PCR mutagenesis of both halves of this palindrome indicates that the TTGT and ACAA motifs are necessary both for normal DNA-protein complex formation in vitro and for full DNA damage-mediated induction of recA. Nevertheless, the basal level of recA expression is not increased when both halves of the TTGTN15ACAA sequence are mutagenized. These data suggest that the R. capsulatus recA gene may be regulated by a positive transcription factor which binds to this palindrome.
Fernàndez de Henestrosa
A R
AR
Department of Genetics and Microbiology, Universitat Autónoma of Barcelona, Bellaterra, Spain.
Labazi
M
M
Mar López
M
M
Barbé
J
J
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004249:DNA Damage; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011938:Rec A Recombinases / Q000235:genetics*; D016354:Rhodobacter capsulatus / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D017931:DNA Primers; D004269:DNA, Bacterial; D014157:Transcription Factors; D011938:Rec A Recombinases