Constitutive expression of PcsB suppresses the requirement for the essential VicR (YycF) response regulator in Streptococcus pneumoniae R6.
We report several new findings about the function of the essential VicRK two-component regulatory system (TCS) in the human pathogen Streptococcus pneumoniae. The vicR-encoded response regulator, vicK-encoded histidine kinase and the protein encoded by the downstream vicX gene are the homologues of the YycF, YycG and YycJ proteins, respectively, studied previously in Bacillus subtilis and Staphylococcus aureus. Using a regulatable promoter, we demonstrated that the VicK histidine kinase is conditionally required for growth of S. pneumoniae. Likewise, we found that the VicX protein is also conditionally required for growth and probably plays a role in the essential signal transduction pathway mediated by VicR and VicK. Recovery of limited substitutions in the conserved aspartate 52 residue (D52) of VicR was consistent with a requirement for phosphorylation of VicR for growth under some conditions. We applied microarrays to characterize the changes in transcription patterns in bacteria depleted for vicRKX operon expression. Our results suggest that the pcsB gene is a target of the VicRK TCS. We present evidence that downregulation of pcsB could account for many of the defects in cell growth, shape, size and morphology observed in bacteria depleted for vicRKX expression. Furthermore, constitutive expression of pcsB+ suppressed the essential requirement for the VicRK TCS and allowed the isolation of vicR null mutants.
Ng
Wai-Leung
WL
Department of Biology, Indiana University, Jordan Hall 142, Bloomington, IN 47405, USA.
Robertson
Gregory T
GT
Kazmierczak
Krystyna M
KM
Zhao
Jingyong
J
Gilmour
Raymond
R
Winkler
Malcolm E
ME
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D000071677:Histidine Kinase; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D009876:Operon; D017354:Point Mutation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism / Q000648:ultrastructure
D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C422465:PcsB protein, Streptococcus agalactiae; C119939:YycF protein, Bacteria; D011494:Protein Kinases; D000071677:Histidine Kinase