Summary
Sporulation of the standard strain of Bacillus subtilis can be induced by decoyinine (a specific inhibitor of guanosine monophosphate synthesis) in the presence of excess ammonium ions, rapidly utilizable carbohydrates and phosphate. Certain developmental mutants unable to sporulate in usual sporulation media can also be induced to sporulate under these conditions. Among the inducible strains are mutants blocked in the citrate cycle, in phosphoenolpyruvate carboxykinase or one of the glycerolphosphate dehydrogenases (NAD dependent or independent). A mutant lacking phosphoglycerate kinase activity and able to grow on glucose and L-malate but not on either carbon source alone, can also be induced to sporulate in a medium containing both glucose and malate. Among other sporulation mutants whose biochemical deficiency is not known, some are weakly inducible whereas many, blocked at sporulation stages O, II, III, or IV, are not inducible. The results show that the functions of the citrate cycle (used for ATP synthesis) and of gluconeogenesis (used for glycerol phosphate and D-glucosamine-6-phosphate synthesis), normally needed for sporulation under conditions of carbohydrate deprivation, are no longer required when sporulation is induced by decoyinine in the presence of carbohydrates. Mutants in which sporulation is highly inducible can be used as a sensitive tool for the detection of other inducing compounds because their spontaneous (uninduced) background of spores is very low.
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Diesterhaft, M.D., Freese, E.: Role of pyruvate carboxylase, phosphoenol-pyruvate carboxykinase, and malic enzyme during growth and sporulation of Bacillus subtilis. J. Biol. Chem. 248, 6062–6070 (1973)
Flechtner, V.R., Hanson, R.S.: Coarse and fine control of citrate synthase from Bacillus subtilis. Biochim. Biophys. Acta 184, 252–262 (1969)
Fortnagel, P., Freese, E.: Analysis of sporulation mutants, II Mutants blocked in the citric acid cycle. J. Bacteriol. 95, 1431–1438 (1968)
Fortnagel, P., Lopez, J.: The regulation of malate dehydrogenase in sporulation mutants of Bacillus subtilis blocked in the citric acid cycle. Biochem. Biophys. Res. Commun. 227, 320–328 (1971)
Freese, E.: Metabolic control of sporulation. In: Spore research (A.N. Barker, G.W. Gould and J. Wolf, eds.) p. 1–32. London: Academic Press 1976
Freese, E., Fortnagel, P.: Analysis of sporulation mutants I. Response of uracil incorporation to carbon sources and other mutant properties. J. Bacteriol. 94, 1957–1969 (1967)
Freese, E.B., Freese, E.: The influence of the developing bacterial spore on the mother cell. Dev. Biol. 60, 453–462 (1977)
Freese, E., Ichikawa, T., Oh, Y.K., Freese, E.B., Prasad, C.: Deficiences or excesses of metabolites interfering with differentiation. Proc. Natl. Acad. Sci. U.S.A. 71, 4188–4193 (1974)
Freese, E., Oh, Y.K., Freese, E.B., Diesterhaft, M.D., Prasad, C.: Suppression of Sporulation of Bacillus subtilis. In: Spores V (H.O. Halvorson, R. Hanson, and L.L. Campbell, eds.) p. 212–221. Washington, D.C.: American Society for Microbiology 1972
Freese, E.B., Cole, R.M., Klofat, W., Freese, E.: Growth, sporulation, and enzyme defects of glucosamine mutants of Bacillus subtilis. J. Bacteriol. 101, 1046–1062 (1970)
Heinze, J.E., Mitani, T., Rich, K.E., Freese, E.: Induction of sporulation by inhibitory purines and related compounds. Biochim. Biophys. Acta 521, 16–26 (1978)
Hoch, J.A.: Genetics of bacterial sporulation. Advances in Genetics 18, 69–98 (1976)
Klofat, W., Picciolo, G., Chapelle, E.W., Freese, E.: Production of adenosine triphosphate in normal cells and sporulation mutants of Bacillus subtilis. J. Biol. Chem. 244, 3270–3276 (1969)
Mitani, T., Heinze, J.E., Freese, E.: Induction of sporulation in Bacillus subtilis by decoyinine or hadacidin. Biochem. Biophys. Res. Commun. 77, 1118–1125 (1977)
Ohne, M., Rutberg, B.: Repression of sporulation in Bacillus subtilis by L-malate. J. Bacteriol. 125, 453–460 (1976)
Ramaley, R.F., Bernlohr, R.W.: Post logarithmic phase metabolism of sporulating microorganisms. J. Biol. Chem. 241, 620–623 (1966)
Rouyard, J.F., Jonesco, H., Schaeffer, P.: Classification génétique de certains mutants de sporulation de Bacillus subtilis Marburg. Ann. Inst. Pasteur (Paris) 113, 675–683 (1967)
Sonenshein, A.L., Cami, B., Brever, J., Cote, R.: Isolation and characterization of rifampin-resistant and streptolydicin-resistant mutants of Bacillus subtilis with altered sporulation properties. J. Bacteriol. 120, 253–265 (1974)
Speck, E.L., Freese, E.: Control of metabolite secretion in Bacillus subtilis. J. Gen. Microbiol. 78, 261–275 (1973)
Sugae, K., Freese, E.: Requirement of acetate and glycine (or serine) for sporulation of Bacillus subtilis. J. Bacteriol. 104, 1074–1085 (1970)
Sumida-Yasumoto, C., Doi, R.H.: Bacillus subtilis ribonucleic acid polymerase mutants conditionally temperative sensitive at various stages of sporulation. J. Bacteriol. 129, 433–444 (1977)
Suhadolnik, R.J.: In: Nucleoside Antibiotics. Ketohexose nucleosides (R.J. Suhadolnik, ed.) p. 96–122. New York, John Wiley and Son 1970
Youston, A.A., Hanson, R.S.: Sporulation of tricarboxylic acid cycle mutants of Bacillus subtilis. J. Bacteriol. 109, 886–894 (1972)
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Communicated by H.G. Wittmann
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Freese, E.B., Vasantha, N. & Freese, E. Induction of sporulation in developmental mutants of Bacillus subtilis . Molec. Gen. Genet. 170, 67–74 (1979). https://doi.org/10.1007/BF00268581
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DOI: https://doi.org/10.1007/BF00268581