Characterization of Bacillus subtilis mutants with a temperature-sensitive intracellular protease.
A colony screening procedure was devised to detect Bacillus subtilis mutants containing temperature-sensitive trypsin-like intracellular protease activity. The enzyme was characterized as a non-sulfhydryl serine protease on the basis of inhibitor studies. It was also inhibited by D- or L-histidine but not by any other amino acid tested. The long-term survival at 45 degrees C of these mutants in a minimal salts medium was decreased, with rapid lysis occurring within 24 h. A D-histidine function in long-term survival and inhibition accounted for the presence of additional protease mutants among survivors of histidine auxotrophs selected for their ability to utilize D-histidine. In addition to being lysed when incubated at 45 degrees C under nongrowth conditions, all of the protease mutants had a decreased rate of protein turnover and produced spores deficient in a major low-molecular-weight spore coat polypeptide. The morphology of the undercoat layers was altered, but there was no effect on spore heat resistance or on germination. The missing spore coat polypeptide appeared to be processed from a larger precursor by cleavage to produce N-terminal histidine. A defect in this protease could account for the lack of processing and thus the absence of this polypeptide in spore coats.
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D001433:Bacteriolysis; D010450:Endopeptidases / Q000378:metabolism*; D006639:Histidine / Q000494:pharmacology; D009154:Mutation; D012697:Serine Endopeptidases; D012808:Sigma Factor; D013171:Spores, Bacterial / Q000032:analysis / Q000502:physiology; D013696:Temperature; D014157:Transcription Factors
D001426:Bacterial Proteins; D012808:Sigma Factor; D014157:Transcription Factors; C092468:spoIIR protein, Bacillus subtilis; C030069:spore-specific proteins, Bacillus; D006639:Histidine; D010450:Endopeptidases; D012697:Serine Endopeptidases