Mutational analysis and membrane topology of ComP, a quorum-sensing histidine kinase of Bacillus subtilis controlling competence development.
ComP is a sensor histidine kinase of Bacillus subtilis required for the signal transduction pathway that initiates the development of competence for genetic transformation. It is believed that ComP senses the presence of ComX, a modified extracellular peptide pheromone, and donates a phosphate to ComA, thereby activating this transcription factor for binding to the srfA promoter. In the present study, fusions to the Escherichia coli proteins PhoA and LacZ and analysis of its susceptibility to the protease kallikrein were used to probe the membrane topology of ComP. These data suggest that ComP contains six or eight membrane-spanning segments and two large extracytoplasmic loops in its N-terminal membrane-associated domain. Deletions were introduced involving the large extracellular loops to explore the role of the N-terminal domain of ComP in signal transduction. The absence of the second loop conferred a phenotype in which ComP was active in the absence of ComX. The implications of these data are discussed.
Public Health Research Institute, New York, New York 10016, USA.
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D000469:Alkaline Phosphatase / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D005838:Genotype; D000071677:Histidine Kinase; D007610:Kallikreins / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017384:Sequence Deletion; D001616:beta-Galactosidase / Q000378:metabolism
D001426:Bacterial Proteins; C087040:ComX protein, Bacillus subtilis; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D000071677:Histidine Kinase; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase; D007610:Kallikreins