Identification of the sequences recognized by the Bacillus subtilis response regulator YclJ
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The Bacillus subtilis yclJ gene encodes an OmpR-type response regulator of a two-component regulatory system with unknown function. A previous DNA microarray experiment suggested that multicopy yclJ greatly enhances the expression of several operons in a cognate kinase (YclK)-deficient strain. To confirm this, lacZ fusion analysis was performed in the yclK background with overexpressed yclJ. As a result, yclHI, ykcBC, and yngABC were indeed positively regulated by YclJ. Gel retardation and DNase I footprint analyses revealed that YclJ binds to the promoter regions of yclHI, ykcBC, and yngABC. Nucleotide sequence analysis of the binding regions suggested that YclJ recognizes a direct repeat of the consensus sequence TTCATANTTT, the upstream half of which has close similarity to the consensus binding sequence of the other OmpR family response regulator PhoP. LacZ fusion analysis of the control region of yngA with deletion or point mutation confirmed that the YclJ-binding sequence is required for the YclJ-mediated activation of yngA. Furthermore, we identified two more YclJ-regulated genes, yycA and yfjR, using bioinformatic analysis of the B. subtilis genome, and it was shown that YclJ binds to those promoters and controls the expression of those genes.
KeywordsYclJ DNA-binding Response regulator
This work was supported by a Grant-in-aid for Scientific Research on Priority Areas (C) “Genome Biology” to T. T., and a Grant-in-aid for Scientific Research (C) to M. O., and (B) to T. T., from the Ministry of Education, Culture, Sports, Science and Technology of Japan. We thank K. Kobayashi for a bacterial strain. We also thank N. Takahashi, T. Furukawa, K. Yamashita, T. Fujio, H. Nishida, Y. Hanawa, and M. Tanaka for technical assistance.
- Hulett FM (2002) The PhoP regulon. In: Sonenshine AL, Hoch JA, Losick R (eds) Bacillus subtilis and its closest relatives: from genes to cells. ASM Press, Washington, DC, pp 193–202Google Scholar
- Huyen NTT, Eiamphungporn W, Mader U, Liebeke M, Lalk M, Hecker M, Helmann JD, Antelmann H (2009) Genome-wide responses to carbonyl electrophiles in Bacillus subtilis: control of the thiol-dependent formaldehyde dehydrogenase Adha and cysteine proteinase YraA by the MerR-family regulator YraB (AdhR). Mol Microbiol 71:876–894CrossRefGoogle Scholar
- Yoshida K, Kobayashi K, Miwa Y, Kang CM, Matsunaga M, Yamaguchi H, Tojo S, Yamamoto M, Nishi R, Ogasawara N, Nakayama T, Fujita Y (2001) Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis. Nucleic Acids Res 29:683–692CrossRefPubMedGoogle Scholar