Isolation and characterization of Bacillus subtilis sigB operon mutations that suppress the loss of the negative regulator RsbX.

Abstract

sigmaB, a transcription factor that controls the Bacillus subtilis general stress response regulon, is activated by either a drop in intracellular ATP or exposure to environmental stress. RsbX, one of seven sigmaB regulators (Rsb proteins) whose genes are cotranscribed with sigmaB, is a negative regulator in the stress-dependent activation pathway. To better define the interactions that take place among the Rsb proteins, we analyzed sigB operon mutations which suppress the high-level sigmaB activity that normally accompanies the loss of RsbX. Each of these mutations was in one of three genes (rsbT, -U, and -V) which encode positive regulators of sigmaB, and they all defined amino acid changes which either compromised the activities of the mutant Rsbs or affected their ability to accumulate. sigmaB activity remained inducible by ethanol in several of the RsbX- suppressor strains. This finding supports the notion that RsbX is not needed as the target for sigmaB activation by at least some stresses. sigmaB activity in several RsbX- strains with suppressor mutations in rsbT or -U was high during growth and underwent a continued, rather than a transient, increase following stress. Thus, RsbX is likely responsible for maintaining low sigmaB activity during balanced growth and for reestablishing sigmaB activity at prestress levels following induction. Although RsbX likely participates in limiting the sigmaB induction response, a second mechanism for curtailing unrestricted sigmaB activation was suggested by the sigmaB induction profile in two suppressor strains with mutations in rsbV. sigmaB activity in these mutants was stress inducible but transient, even in the absence of RsbX.