Genome-wide identification of genes directly regulated by the pleiotropic transcription factor Spx in Bacillus subtilis.
The transcriptional regulator Spx plays a key role in maintaining the redox homeostasis of Bacillus subtilis cells exposed to disulfide stress. Defects in Spx were previously shown to lead to differential expression of numerous genes but direct and indirect regulatory effects could not be distinguished. Here we identified 283 discrete chromosomal sites potentially bound by the Spx-RNA polymerase (Spx-RNAP) complex using chromatin immunoprecipitation of Spx. Three quarters of these sites were located near Sigma(A)-dependent promoters, and upon diamide treatment, the fraction of the Spx-RNAP complex increased in parallel with the number and occupancy of DNA sites. Correlation of Spx-RNAP-binding sites with gene differential expression in wild-type and Δspx strains exposed or not to diamide revealed that 144 transcription units comprising 275 genes were potentially under direct Spx regulation. Spx-controlled promoters exhibited an extended -35 box in which nucleotide composition at the -43/-44 positions strongly correlated with observed activation. In vitro transcription confirmed activation by oxidized Spx of seven newly identified promoters, of which one was also activated by reduced Spx. Our study globally characterized the Spx regulatory network, revealing its role in the basal expression of some genes and its complex interplay with other stress responses.
INRA, UMR 1319 Micalis, France.
Tatiana
T
Rochat
Pierre
P
Nicolas
Olivier
O
Delumeau
Alžbeta
A
Rabatinová
Jana
J
Korelusová
Aurélie
A
Leduc
Philippe
P
Bessières
Etienne
E
Dervyn
Libor
L
Krásny
Philippe
P
Noirot
D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003958:Diamide / Q000633:toxicity; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D011401:Promoter Regions, Genetic; D018085:Regulon; D013312:Stress, Physiological / Q000235:genetics; D013439:Sulfhydryl Reagents / Q000633:toxicity; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D013439:Sulfhydryl Reagents; D014157:Transcription Factors; D003958:Diamide; D012321:DNA-Directed RNA Polymerases