The H2O2 stress-responsive regulator PerR positively regulates srfA expression in Bacillus subtilis.
srfA is an operon required for the synthesis of surfactin and the development of genetic competence in Bacillus subtilis. We observed that the expression of srfA is downregulated upon treatment with H2O2. Thus, we examined the involvement of several oxidative stress-responsive transcription factors in srfA expression. Our DNA microarray analysis revealed that the H2O2 stress-responsive regulator PerR is required for srfA expression. This was confirmed by lacZ fusion analysis. A ComX feeding assay and epistatic analyses revealed that the role of PerR in srfA expression is independent of other known regulators of srfA expression, namely, comQXP, rapC, and spx. Gel mobility shift and footprint assays revealed that PerR binds directly to two tandemly arranged noncanonical PerR boxes located in the upstream promoter region of srfA. A transcriptional srfA-lacZ fusion lacking both PerR boxes showed diminished and PerR-independent expression, indicating that the PerR boxes we identified function as positive cis elements for srfA transcription.
Hayashi
Kentaro
K
Department of Marine Science, School of Marine Science and Technology, Tokai University, 3-20-1 Orido, Shimizu, Shizuoka 424-8610, Japan.
Ohsawa
Taku
T
Kobayashi
Kazuo
K
Ogasawara
Naotake
N
Ogura
Mitsuo
M
D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D004950:Esterases / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D005816:Genetic Complementation Test; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007763:Lac Operon; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000502:physiology*; D010453:Peptide Synthases / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology; D012097:Repressor Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology; D014166:Transferases / Q000378:metabolism
D001426:Bacterial Proteins; C087040:ComX protein, Bacillus subtilis; D008565:Membrane Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C122138:peroxide repressor proteins; C070739:comQ protein, Bacillus subtilis; D006861:Hydrogen Peroxide; D014166:Transferases; C064995:comP protein, Bacillus subtilis; D004950:Esterases; C101114:RapC protein, Bacillus subtilis; D010453:Peptide Synthases; C069542:surfactin synthetase