Involvement of Clp protease activity in modulating the Bacillus subtilissigmaw stress response.
The induction of Bacillus subtilis genes controlled by the extracytoplasmic function alternative sigma factor sigmaW is strongly impaired in a strain deleted for the ClpP peptidase gene and in a double knockout of the ClpX and ClpE ATPase genes. Truncated soluble forms of the sigmaW anti-sigma factor RsiW are stabilized in a clpP minus strain as revealed by the green fluorescent reporter protein fused to the N-terminus of RsiW and by pulse-chase experiments. Conserved alanine residues are present in the transmembrane region of RsiW, and mutations in these positions abolish induction of sigmaW-controlled genes. Following alkaline shock, a truncated cytoplasmic form of RsiW is detectable in a strain expressing a triple alanine mutant allele of rsiW. These data point to a mechanism where the trans-membrane segment of RsiW contains a cryptic proteolytic tag that is uncovered as a result of intramembrane proteolysis of RsiW by RasP (YluC). After RasP-clipped RsiW is detached from the membrane, this proteolytic tag becomes crucial for the complete degradation of RsiW by cytoplasmic proteases and the release of sigmaW. ClpXP plays a major role in this third proteolytic step of stress-induced degradation of RsiW. Overexpression of SsrA-tagged green fluorescent protein as a ClpXP substrate protein reduces alkali induction of a sigmaW-controlled gene by a factor of about three, indicating that a titration mechanism is able to tune the sigmaW-mediated stress response to the cellular state.
Zellmeier
Stephan
S
Institute of Genetics, University of Bayreuth, D-95440 Bayreuth, Germany.
Schumann
Wolfgang
W
Wiegert
Thomas
T
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000409:Alanine / Q000737:chemistry / Q000235:genetics; D000468:Alkalies / Q000494:pharmacology; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002462:Cell Membrane; D017124:Conserved Sequence; D049071:Endopeptidase Clp / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000235:genetics; D018085:Regulon / Q000235:genetics; D012808:Sigma Factor / Q000235:genetics* / Q000378:metabolism*
D000468:Alkalies; D001426:Bacterial Proteins; C413458:SigW protein, Bacillus subtilis; D012808:Sigma Factor; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases; D000409:Alanine