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Microbiology 153 (2007), 667-675; DOI  10.1099/mic.0.2006/003673-0
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Microbiology 153 (2007), 667-675; DOI  10.1099/mic.0.2006/003673-0
© 2007 Society for General Microbiology

The Bacillus subtilis NatK–NatR two-component system regulates expression of the natAB operon encoding an ABC transporter for sodium ion extrusion

Mitsuo Ogura, Kensuke Tsukahara, Kentaro Hayashi and Teruo Tanaka

Institute of Oceanic Research and Development, Tokai University, 3-20-1 Orido-Shimizu, Shizuoka 424-8610, Japan

Correspondence
Mitsuo Ogura
oguram{at}scc.u-tokai.ac.jp

A previous microarray analysis suggested that multicopy yccH, encoding a function-unknown response regulator, enhances expression of natAB, which encodes a two-gene ATP-binding cassette transporter involved in the extrusion of sodium ions. The two-component regulatory system YccG–YccH was therefore renamed NatK–NatR. Here, this observation was confirmed by a lacZ fusion analysis using a strain carrying natA–lacZ. Further, in both natK and natR mutants, natA–lacZ expression was completely abolished, indicating that the NatK–NatR system positively regulates the expression of natAB. In a gel retardation analysis, NatR bound to the natA promoter region. Using purified His-tagged NatR, DNase I footprinting analysis of the natA promoter region suggested that a direct repeat of [TTCA(G)CGACA], separated by a 12 bp space, would be recognized by NatR. Deleted and mutagenized promoter regions of natA were analysed using a lacZ fusion, and it was confirmed that the direct repeat is critical for natA activation by NatR.

Abbreviations: ABC, ATP-binding cassette

A supplementary figure showing the structure of plasmid pDG-N17 is available with the online version of this paper.






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