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The H₂O₂ Stress-Responsive Regulator PerR Positively Regulates srfA Expression in Bacillus subtilis

Kentaro Hayashi,^1, Taku Ohsawa,^1, Kazuo Kobayashi,² Naotake Ogasawara,² and Mitsuo Ogura^1*

Department of Marine Science, School of Marine Science and Technology, Tokai University, 3-20-1 Orido, Shimizu, Shizuoka 424-8610, Japan,¹ Graduate School of Information Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-019, Japan²

Received 3 June 2005/ Accepted 22 July 2005

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.

These authors contributed equally to this work.

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