Secreted-protein response to sigmaU activity in Streptomyces coelicolor.
The filamentous bacterium Streptomyces coelicolor forms an aerial mycelium as a prerequisite to sporulation, which occurs in the aerial hyphae. Uncontrolled activity of the extracytoplasmic function sigma factor sigmaU blocks the process of aerial mycelium formation in this organism. Using a green fluorescent protein transcriptional reporter, we have demonstrated that sigU transcription is autoregulated. We have defined a sigmaU-dependent promoter sequence and used this to identify 22 likely sigmaU regulon members in the S. coelicolor genome. Since many of these genes encode probable secreted proteins, we characterized the extracellular proteome of a mutant with high sigmaU activity caused by disruption of rsuA, the presumed cognate anti-sigma factor of sigmaU. This mutant secreted a much greater quantity and diversity of proteins than the wild-type strain. Peptide mass fingerprinting was used to identify 79 proteins from the rsuA mutant culture supernatant. The most abundant species, SCO2217, SCO0930, and SCO2207, corresponded to secreted proteins or lipoproteins of unknown functions whose genes are in the proposed sigmaU regulon. Several unique proteases were also detected in the extracellular proteome of the mutant, and the levels of the protease inhibitor SCO0762 were much reduced compared to those of the wild type. Consequently, extracellular protease activity was elevated about fourfold in the rsuA mutant. The functions of the proteins secreted as a result of sigmaU activity may be important for combating cell envelope stress and modulating morphological differentiation in S. coelicolor.
Department of Chemistry, Williams College, Williamstown, Massachusetts 01267, USA.
Nadria D
ND
Gordon
Geri L
GL
Ottaviano
Sarah E
SE
Connell
Gregory V
GV
Tobkin
Crystal H
CH
Son
Sebastian
S
Shterental
Amy M
AM
Gehring
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019751:Intramolecular Transferases / Q000235:genetics; D010449:Peptide Mapping; D011401:Promoter Regions, Genetic; D020543:Proteome; D012808:Sigma Factor / Q000378:metabolism*; D048372:Streptomyces coelicolor / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D020543:Proteome; D012808:Sigma Factor; D049452:Green Fluorescent Proteins; D019751:Intramolecular Transferases; C094732:16S RNA pseudouridine 516 synthase, E coli