Temporal regulation of σ^B by partner-switching mechanism at a distinct growth stage in Bacillus cereus

The alternative transcription factor σ^B in Bacillus cereus governs the transcription of a number of genes that confer protection against general stress. This transcription factor is regulated by protein-protein interactions among RsbV, RsbW, σ^B, RsbY, RsbM and RsbK, all encoded in the sigB cluster. Among these regulatory proteins, RsbV, RsbW and σ^B comprise a partner-switching mechanism. Under normal conditions, σ^B remains inactive by associating with anti-sigma factor RsbW, which prevents σ^B from binding to the core RNA polymerase. During environmental stress, RsbK activates RsbY to hydrolyze phosphorylated RsbV, and the dephosphorylated RsbV then sequesters RsbW to liberate σ^B from RsbW. Although the σ^B partner-switching module is thought to be the core mechanism for σ^B regulation, the actual protein-protein interactions among these three proteins in the cell remain to be investigated. In the current study, we show that RsbW and RsbV form a long-lived complex under transient stress treatment, resulting in high persistent expression of RsbV, RsbW and σ^B from mid-log phase to stationary phase. Full sequestration of RsbW by excess RsbV and increased RsbW:RsbV complex stability afforded by cellular ADP contribute to the prolonged activation of σ^B. Interestingly, the high expression levels of RsbV, RsbW and σ^B were dramatically decreased beginning from the transition stage to the stationary phase. Thus, protein interactions among σ^B partner-switching components are required for the continued induction of σ^B during environmental stress in the log phase and significant down-regulation of σ^B is observed in the stationary phase. Our data show that σ^B is temporally regulated in B. cereus.