Summary: The nucleotide sequence of wprA, a protease-encoding gene of Bacillus subtilis 168, is reported. The gene, expressed during the exponential growth phase, belongs to a monocistronic operon. WprA is a 96 kDa polypeptide endowed with a signal peptide, as well as a propeptide. Upon processing and export, it gives rise to two previously identified cell-wall-bound proteins, CWBP23 and 52. Processing of WprA exhibits a novel feature of protein export, whereby removal of the middle part of the molecule accompanies the targeting to the cell wall of its N- and C-terminal parts, which correspond to CWBP23 and 52, respectively. Sequence analyses and enzymic assays reveal that CWBP52 is a serine protease. Growth rate, cell morphology, sporulation and motility of wprA mutants apparently do not differ from those of the parent strain.
AnagnostopoulosC., PiggotP. J., HochJ. A.1993; The genetic map of Bacillus subtilis. In Bacillus subtilis and Other Grampositive bacteria pp. 425–461 Edited by SonensheinA. L., HochJ. A., LosickR. Washington, DC:: American Society for Microbiology;
ChungC. T., NiemelaS. L., MillerR. H.1989; One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA862172–2175
CondonC., PutzerH., Grunberg-ManagoM.1996; Processing of the leader mRNA plays a major role in the induction of expression following threonine starvation in Bacillus subtilis. Proc Natl Acad Sci USA936992–6997
FosterS. J.1991; Cloning, expression, sequence analysis and biochemical characterization of an autolytic amidase of Bacillus subtilis 168 trpC2. J Gen Microbiol 137:1987–1998
FosterS. J.1993; Molecular analysis of three major wall- associated proteins of Bacillus subtilis 168: evidence for processing of the product of a gene encoding a 258 kDa precursor two-domain ligand-binding protein. Mol Microbiol 8:299–310
JacobsM., EliassonM., UhlenM., FlockJ. I.1985; Cloning, sequencing and expression of subtilisin Carlsberg from Bacillus licheniformis. Nucleic Acids Res 13:8913–8926
KaramataD., GrossJ. D.1970; Isolation and genetic analysis of temperature-sensitive mutants of B. subtilis defective in DNA synthesis. Mol Gen Genet 108:277–287
KoideY., NakamuraA., UozumiT., BeppuT.1986; Cloning and sequencing of the major intracellular serine protease gene of Bacillus subtilis. J Bacteriol 167:110–116
LazarevicV., MargotP., SoldoB., KaramataD.1992; Sequencing and analysis of the Bacillus subtilis lytRABC divergon: a regulatory unit encompassing the structural genes of the N-acetylmuramoyl-L-alanine amidase and its modifier. J Gen Microbiol 138:1949–1961
LongchampP. F., MauélC., KaramataD.1994; Lytic enzymes associated with defective prophages of Bacillus subtilis: sequencing and characterization of the region comprising the N-acetyl- muramoyl-L-alanine amidase gene of prophage PBSX. Microbiology 140:1855–1867
MargotP., MauélC., KaramataD.1994; The gene of the N-acetylglucosaminidase, a Bacillus subtilis 168 cell wall hydrolase not involved in vegetative cell autolysis. Mol Microbiol 12:535–545
MauélC., YoungM., MargotP., KaramataD.1989; The essential nature of teichoic acids in Bacillus subtilis as revealed by insertional mutagenesis. Mol Gen Genet 215:388–394
MelounB., BaudysM., KostkaV., HausdorfG., FrommelG., HohnemW. E.1985; Complete primary structure of thermitase from Thermoactinomyces vulgaris and its structural features related to the subtilisin-type proteinase. FEBS Lett 183:195–200
MonodM.1992; Isolation of Aspergillus fumigatus genes using oligonucleotide probes. In Molecular Biology of Pathogenic Fungi. Laboratory Manual pp. 33–40 Edited by MarescaB., KobayashiG. S. New York:: Telos Press;
OhtaY., HojoH., AimotoS., KobayashiT., ZhuX., JordanF., InouyeM.1991; Pro-peptide as an intermolecular chaperone: renaturation of denatured subtilisin E with a synthetic pro-peptide. Mol Microbiol 5:1507–1510
OultramJ. D., PeckH., BrehmJ. K., ThompsonD., SwinfielT. J., MintonN. P.1988; Introduction of genes for leucine biosynthesis from Clostridium pasteurianum into Clostridium aceto-butylicum. Mol Gen Genet 214:177–179
PeroJ., SlomaA.1993; Proteases. In Bacillus subtilis and Other Gram-Positive Bacteria pp. 939–952 Edited by SonensheinA. L., HochJ. A., LosickR. Washington, DC:: American Society for Microbiology;
PollockM. R.1965; Purification and properties of penicillinases from two strains of Bacillus licheniformis: a chemical, physiochemical and physiological comparison. Biochemistry 94:666–675
RotenC. -A., PagniM., MargotP., TouriF., KaramataD.1994; Specific labeling of diaminopimelate: a radioassay for the determination of the peptidoglycan cross-linking index. Anal Biochem 223:208–211
SchlaeppiJ. M., PooleyH. M., KaramataD.1982; Identification of cell wall subunits in Bacillus subtilis and analysis of their segregation during growth. J Bacteriol 149:329–337
SchneewindO., JonesK. F., FischettiV. A.1990; Sequence and structural characteristics of the trypsin-resistant T6 surface protein of group A streptococci. J Bacteriol 172:3310–3317
SekiguchiJ., AkeoK., YamamotoH., KhasanovF. K., AlonsoJ. C., KurodaA.1995; Nucleotide sequence and regulation of a new putative cell wall hydrolase gene, cwlD, which affects germination in Bacillus subtilis. J Bacteriol 177:5582–5589
SerrorP., AzevedoV., EhrlichS. D.1993; An ordered collection of Bacillus subtilis DNA segments in yeast artificial chromosomes. In Bacillus subtilis and Other Gram-positive Bacteria pp. 473–474 Edited by SonensheinA. L., HochJ. A., LosickR. Washington, DC:: American Society for Microbiology;
SmithT. J., FosterS. J.1995; Characterization of the involvement of two compensatory autolysins in mother cell lysis during sporulation of Bacillus subtilis168. J Bacteriol 177:3855–3862
StahlM. L., FerrariE.1984; Replacement of the Bacillus subtilissubtilisin structural gene with an in vitro-derived deletion mutation. J Bacteriol 158:411–418
StuderR. E.1988Caracterisation de la paroi native de Bacillus subtilis etude des proteines qui lui sont associees These de doctorat, Universite de Lausanne.
Cell wall proteins in Bacillus subtilis. In Antibiotic Inhibition of Bacterial Cell Surface Assembb and Fgnction pp. 146–150 Edited by ActorP., Daneo-MooreL., HigginsM. L., SaltonM. R. G., ShockmanG. D. Washington, DC:: American Society for Microbiology;