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. 1973 May;114(2):517–527. doi: 10.1128/jb.114.2.517-527.1973

Inactivation of Aspartic Transcarbamylase in Sporulating Bacillus subtilis: Demonstration of a Requirement for Metabolic Energy

Louise M Waindle 1, R L Switzer 1
PMCID: PMC251804  PMID: 4196242

Abstract

The aspartic transcarbamylase (ATCase) activity of Bacillus subtilis cells disappears rapidly from stationary-phase cells prior to sporulation. ATCase activity does not appear in the culture fluid during the stationary phase; hence the enzyme appears to be inactivated in the cells. The enzyme is inactivated normally in two different mutants lacking proteases; the activity is very stable in crude extracts of cells or in the culture fluid. These results suggest that ATCase is not inactivated by the general proteolysis that occurs in sporulating bacteria. The inactivation of ATCase can be completely inhibited after it has begun by oxygen starvation or addition of fluoroacetate. Inhibitors of oxidative phosphorylation and electron transport also interrupt the inactivation of ATCase. The inactivation of ATCase is very slow in two mutant strains that are deficient in enzymes of tricarboxylic acid cycle. Addition of gluconate to stationary cultures of the mutant strains, which is known to restore depleted adenosine 5′-triphosphate pools in these bacteria, also restores inactivation of ATCase. These experiments support the conclusion that the generation of metabolic energy is necessary for the inactivation of ATCase in stationary cells. ATCase activity is stable in growing cells in which ATCase synthesis is repressed by addition of uracil; the enzyme is inactivated normally, however, when such cells cease growing.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Carls R. A., Hanson R. S. Isolation and characterization of tricarboxylic acid cycle mutants of Bacillus subtilis. J Bacteriol. 1971 Jun;106(3):848–855. doi: 10.1128/jb.106.3.848-855.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chow C. T., Takahashi I. Acid-soluble nucleotides in an asporogenous mutant of Bacillus subtilis. J Bacteriol. 1972 Mar;109(3):1175–1180. doi: 10.1128/jb.109.3.1175-1180.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cox D. P., Hanson R. S. Catabolite repression of aconitate hydratase in Bacillus subtilis. Biochim Biophys Acta. 1968 Apr 16;158(1):36–44. doi: 10.1016/0304-4165(68)90069-x. [DOI] [PubMed] [Google Scholar]
  4. Deutscher M. P., Kornberg A. Biochemical studies of bacterial sporulation and germination. 8. Patterns of enzyme development during growth and sporulation of Baccillus subtilis. J Biol Chem. 1968 Sep 25;243(18):4653–4660. [PubMed] [Google Scholar]
  5. GOLLAKOTA K. G., HALVORSON H. O. BIOCHEMICAL CHANGES OCCURRING DURING SPORULATION OF BACILLUS CEREUS T. II. EFFECT OF ESTERS OF ORGANIC ACIDS ON SPORULATION. J Bacteriol. 1963 Jun;85:1386–1393. doi: 10.1128/jb.85.6.1386-1393.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gallant J., Margason G. Amino acid control of messenger ribonucleic acid synthesis in Bacillus subtilis. J Biol Chem. 1972 Apr 25;247(8):2289–2294. [PubMed] [Google Scholar]
  7. Goldberg A. L. Degradation of abnormal proteins in Escherichia coli (protein breakdown-protein structure-mistranslation-amino acid analogs-puromycin). Proc Natl Acad Sci U S A. 1972 Feb;69(2):422–426. doi: 10.1073/pnas.69.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grandgenett D. P., Stahly D. P. Control of diaminopimelate decarboxylase by L-lysine during growth and sporulation of Bacilluscereus. J Bacteriol. 1971 May;106(2):551–560. doi: 10.1128/jb.106.2.551-560.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gray B. H., Bernlohr R. W. The regulation of aspartokinase in Bacillus licheniformis. Biochim Biophys Acta. 1969 Apr 22;178(2):248–261. doi: 10.1016/0005-2744(69)90394-5. [DOI] [PubMed] [Google Scholar]
  10. HEYTLER P. G. uncoupling of oxidative phosphorylation by carbonyl cyanide phenylhydrazones. I. Some characteristics of m-Cl-CCP action on mitochondria and chloroplasts. Biochemistry. 1963 Mar-Apr;2:357–361. doi: 10.1021/bi00902a031. [DOI] [PubMed] [Google Scholar]
  11. Hageman J. H., Carlton B. C. An enzymatic and immunological comparison of two proteases from a transformable Bacillus subtilis with the "subtilisins". Arch Biochem Biophys. 1970 Jul;139(1):67–79. doi: 10.1016/0003-9861(70)90045-7. [DOI] [PubMed] [Google Scholar]
  12. Holzer H., Duntze W. Metabolic regulation by chemical modification of enzymes. Annu Rev Biochem. 1971;40:345–374. doi: 10.1146/annurev.bi.40.070171.002021. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Leitzmann C., Bernlohr R. W. Threonine dehydratase of Bacillus licheniformis. II. Regulation during development. Biochim Biophys Acta. 1968 Feb 5;151(2):461–472. doi: 10.1016/0005-2744(68)90114-9. [DOI] [PubMed] [Google Scholar]
  15. Mandelstam J., Waites W. M. Sporulation in Bacillus subtilis. The role of exoprotease. Biochem J. 1968 Oct;109(5):793–801. doi: 10.1042/bj1090793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. NAKATA H. M., HALVORSON H. O. Biochemical changes occurring during growth and sporulation of Bacillus cereus. J Bacteriol. 1960 Dec;80:801–810. doi: 10.1128/jb.80.6.801-810.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Porter R. W., Modebe M. O., Stark G. R. Aspartate transcarbamylase. Kinetic studies of the catalytic subunit. J Biol Chem. 1969 Apr 10;244(7):1846–1859. [PubMed] [Google Scholar]
  18. Prescott L. M., Jones M. E. Modified methods for the determination of carbamyl aspartate. Anal Biochem. 1969 Dec;32(3):408–419. doi: 10.1016/s0003-2697(69)80008-4. [DOI] [PubMed] [Google Scholar]
  19. Prestidge L., Gage V., Spizizen J. Protease activities during the course of sporulation on Bacillus subtilis. J Bacteriol. 1971 Sep;107(3):815–823. doi: 10.1128/jb.107.3.815-823.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sadoff H. L., Celikkol E., Engelbrecht H. L. Conversion of bacterial aldolase from vegetative to spore form by a sporulation-specific protease. Proc Natl Acad Sci U S A. 1970 Jul;66(3):844–849. doi: 10.1073/pnas.66.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Setlow P., Kornberg A. Biochemical studies of bacterial sporulation and germination. 23. Nucleotide metabolism during spore germination. J Biol Chem. 1970 Jul 25;245(14):3645–3652. [PubMed] [Google Scholar]
  23. Stahly D. P., Bernlohr R. W. Control of aspartokinase during development of Bacillus licheniformis. Biochim Biophys Acta. 1967;146(2):467–476. doi: 10.1016/0005-2744(67)90230-6. [DOI] [PubMed] [Google Scholar]
  24. Swanton M., Edlin G. Isolation and characterization of an RNA relaxed mutant of B. subtilis. Biochem Biophys Res Commun. 1972 Jan 31;46(2):583–588. doi: 10.1016/s0006-291x(72)80179-7. [DOI] [PubMed] [Google Scholar]
  25. Yousten A. A., Hanson R. S. Sporulation of tricarboxylic acid cycle mutants of Bacillus subtilis. J Bacteriol. 1972 Feb;109(2):886–894. doi: 10.1128/jb.109.2.886-894.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

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