ABSTRACT
Temporal and spatial gene regulation during Bacillus subtilis sporulation involves the activation and inactivation of multiple sigma subunits of RNA polymerase in a cascade. In the mother cell compartment of sporulating cells, expression of thesigE gene, encoding the earlier-acting sigma factor, ςE, is negatively regulated by the later-acting sigma factor, ςK. Here, it is shown that the negative feedback loop does not require SinR, an inhibitor of sigEtranscription. Production of ςK about 1 h earlier than normal does affect Spo0A, which when phosphorylated is an activator of sigE transcription. A mutation in thespo0A gene, which bypasses the phosphorelay leading to the phosphorylation of Spo0A, diminished the negative effect of early ςK production on sigE expression early in sporulation. Also, early production of ςK reduced expression of other Spo0A-dependent genes but not expression of the Spo0A-independent ald gene. In contrast, bothsigE and ald were overexpressed late in development of cells that fail to make ςK. Theald promoter, like the sigE promoter, is believed to be recognized by ςA RNA polymerase, suggesting that ςK may inhibit ςA activity late in sporulation. To exert this negative effect, ςKmust be transcriptionally active. A mutant form of ςKthat associates with core RNA polymerase, but does not direct transcription of a ςK-dependent gene, failed to negatively regulate expression of sigE or aldlate in development. On the other hand, the negative effect of early ςK production on sigE expression early in sporulation did not require transcriptional activity of ςK RNA polymerase. These results demonstrate that ςK can negatively regulate sigE expression by two different mechanisms, one observed when ςK is produced earlier than normal, which does not require ςKto be transcriptionally active and affects Spo0A, and the other observed when ςK is produced at the normal time, which requires ςK RNA polymerase transcriptional activity. The latter mechanism facilitates the switch from ςE to ςK in the cascade controlling mother cell gene expression.
Gene encoding the 37,000-dalton minor sigma factor of Bacillus subtilis RNA polymerase: isolation, nucleotide sequence, chromosomal locus, and cryptic function.
