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Characterization and function of intracellular proteases in sporulating bacillus cereus

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Abstract

Intracellular proteases from sporulating Bacillus cereus have been purified by ammonium sulfate fractionation, heat treatment and DEAE cellulose column chromatography. After the last purification step, two protease activities, with an activity ratio of about thirty to one are resolved. Both proteases are resistant to o-phenanthroline but sensitive to phenyl methyl sulfonyl fluoride. Their separation by polyacrylamide gel electrophoresis and DEAE cellulose column chromatography, their difference in heat sensitivity and a mutation affecting only the major intracellular protease (IP1) suggest that the two are products of distinct genes. An IP1 mutant previously shown to produce coat defective spores (4) also turnsover protein with a reduced rate during late sporulation stages. Correlated with the slower turnover rate in this mutant is the more rapid disappearance of IP1. A partial revertant of this mutant has a protein turnover rate intermediate between that of the original mutant and wild type. These correlations imply that IP1 has an important role in protein turnover during sporulation.

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References

  • Aronson, A. I., Angelo, N., Holt, S. C.: Regulation of extracellular protease production in Bacillus cereus T: characterization of mutants producing altered amounts of protease. J. Bacteriol. 106, 1016–1025 (1971)

    Google Scholar 

  • Aronson, A. I.: The function of various proteases in bacterial sporulation, pp. 33–37. In: Regulation de la sporulation microbienne. Colloq. Int. C.N.R.S. No. 227 (1973)

  • Both, G. W., McInnes, J. L., Hanlon, J. G., May, B. K., Elliott, W. H.: Evidence for an accumulation of messenger RNA specific for extracellular protease and its relevance to the mechanism of enzyme secretion in bacteria. J. Mol. Biol. 67, 199–217 (1972)

    Google Scholar 

  • Cheng, Y. E., Aronson, A. I.: Alterations of spore coat processing and protein turnover in a Bacillus cereus mutant with a defective postexponential intracellular protease. Proc. Natl. Acad. Sci. U.S.A. 74, 1254–1258 (1977)

    Google Scholar 

  • Doi, R. H.: Role of proteases in sporulation. In: Current topics in cellular regulation, Vol. 6 (B. L. Horecker, E. R. Stadtman, eds.), pp. 1–20. New York: Academic Press 1972

    Google Scholar 

  • Hageman, J. H., Carlton, B. C.: Effects of mutational loss of specific intracellular proteases on the sporulation of Bacillus subtilis. J. Bacteriol. 114, 612–617 (1973)

    Google Scholar 

  • Kornberg, A., Spudich, J. A., Nelson, D. L., Deutscher, M. P.: Origin of proteins in sporulation. Ann. Rev. Biochem. 37, 51–78 (1968)

    Google Scholar 

  • Levisohn, S., Aronson, A. I.: Regulation of extracellular protease production in Bacillus cereus. J. Bacteriol. 93, 1023–1030 (1967)

    Google Scholar 

  • Linn, J. G., Greenleaf, A. L., Shorenstein, R. G., Losick, R.: Loss of the sigma activity of RNA polymerase of Bacillus subtilis during sporulation. Proc. Natl. Acad. Sci. U.S.A. 70, 1865–1869 (1973)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Mandelstam, J., Waits, W. M.: Sporulation in B. subtilis. The role of exoprotease. Biochem. J. 109, 793–802 (1968)

    Google Scholar 

  • Michel, J., Millet, J.: Physiological studies on early-blocked sporulation mutants of Bacillus subtilis. J. Appl. Bacteriol. 33, 220–227 (1970)

    Google Scholar 

  • Millet, J.: Caractérisation d'une endopeptidase cytoplasmique chez Bacillus megaterium en voie de sporulation. C. R. Acad Sci. 272, 1806–1809 (1971)

    Google Scholar 

  • Millet, J., Aubert, J. P.: Etude de la megateriopeptidase, protéase exocellulaire de Bacillus megaterium. Biosynthese et role physiologique. Ann. Inst. Pasteur, Paris 117, 461–473 (1969)

    Google Scholar 

  • Millet, J., Larribe, M., Aubert, J. P.: Mutant thermosensible de B. subtilis affecté dans la sporulation et la sérylprotéase extracellaire. Biochimie 58, 109–117 (1976)

    Google Scholar 

  • Reysset, G., Millet, J.: Characterization of an intracellular protease in B. subtilis during sporulation. Biochem. Biophys. Res. Commun. 49, 328–334 (1972)

    Google Scholar 

  • Sadoff, H. L., Celikol, E., Engelbrecht, A. L.: Conversion of bacterial aldolase from vegetative to spore form by a sporulation-specific protease. Proc. Natl. Acad. Sci. U.S.A. 66, 844–849 (1970)

    Google Scholar 

  • Schaeffer, P.: Sporulation and the production of antibiotics, exoenzymes, and exotoxins. Bacteriol. Rev 33, 48–71 (1969)

    Google Scholar 

  • Setlow, P.: Identification and localization of the major proteins degraded during germination of Bacillus megaterium spores. J. Biol. Chem 250, 8159–8167 (1975)

    Google Scholar 

  • Setlow, P.: Purification and properties of a specific proteolytic enzyme present in spores of Bacillus megaterium. J. Biol. Chem. 251, 7853–7862 (1976)

    Google Scholar 

  • Setlow, P., Primus, G.: Protein degradation and amino acid metabolism during germination of Bacillus megaterium spores In: Spores, Vol. VI (P. Gerhardt, R. N. Costilow, H. L. Sadoff, eds.), pp. 451–457 Washington, D. C.: American Society for Microbiology 1975

    Google Scholar 

  • Szulmajster, J., Keryer, E.: Isolation and properties of thermosensitive sporulation mutants of Bacillus subtilis deficient in intracellular protease activity. In: Spores, Vol. VI (P. Gerhardt, R. N. Costilow, H. L. Sadoff, eds.), pp. 271–278 Washington, D.C.: American Society for Microbiology 1975

    Google Scholar 

  • Williams, D. E., Reisfeld, R. A.: Disc electrophoresis in polyacrylamide gels: Extension to new conditions of pH and buffer. Annals N. Y. Acad. Sci. 121, 373–381 (1964)

    Google Scholar 

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Cheng, YS.E., Aronson, A.I. Characterization and function of intracellular proteases in sporulating bacillus cereus . Arch. Microbiol. 115, 61–66 (1977). https://doi.org/10.1007/BF00427846

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