Characterization of a new sigma-K-dependent peptidoglycan hydrolase gene that plays a role in Bacillus subtilis mother cell lysis.
Bacillus subtilis produces a 30-kDa peptidoglycan hydrolase, CwlH, during the late sporulation phase. Disruption of yqeE led to a complete loss of CwlH formation, indicating the identity of yqeE with cwlH. Northern blot analysis of cwlH revealed a 0.8-kb transcript after 6 to 7.5 h for the wild-type strain but not for the sigma(F), sigma(E), sigma(G), and sigma(K) mutants. Expression of the sigma(K)-dependent cwlH gene depended on gerE. Primer extension analysis also suggested that cwlH is transcribed by Esigma(K) RNA polymerase. CwlH produced in Escherichia coli harboring a cwlH plasmid is an N-acetylmuramoyl-L-alanine amidase (EC 3.5.1.28) and exhibited an optimum pH of 7.0 and high-level binding to the B. subtilis cell wall. A cwlC cwlH double mutation led to a lack of mother cell lysis even after 7 days of incubation in DSM medium, but the single mutations led to mother cell lysis after 24 h.
Nugroho
F A
FA
Department of Applied Biology, Faculty of Textile Science, Shinshu University, Ueda-shi, Nagano 386-8567, Japan.
Yamamoto
H
H
Kobayashi
Y
Y
Sekiguchi
J
J
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000166:cytology / Q000502:physiology*; D001426:Bacterial Proteins; D001433:Bacteriolysis / Q000502:physiology*; D001483:Base Sequence; D002473:Cell Wall / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D013171:Spores, Bacterial / Q000502:physiology; D013379:Substrate Specificity; D014157:Transcription Factors / Q000378:metabolism*
D001426:Bacterial Proteins; D014157:Transcription Factors; C084190:sigma K; C122132:CwlH protein, Bacillus subtilis; D009238:N-Acetylmuramoyl-L-alanine Amidase