Abstract
The citB of Bacillus subtilis codes for aconitase (D. W. Dingman and A. L. Sonenshein, J. Bacteriol. 169:3060-3065). By direct measurements of citB mRNA levels and by measurements of beta-galactosidase activity in a strain carrying a citB-lacZ fusion, we have examined the expression of citB during growth and sporulation. When cells were grown in nutrient broth sporulation medium, citB mRNA appeared in mid- to late-exponential phase and disappeared by the second hour of sporulation. This timing corresponded closely to the kinetics of appearance of aconitase enzyme activity. Decoyinine, a compound that induces sporulation in a defined medium, caused a rapid simultaneous increase in aconitase activity and citB transcription. After decoyinine addition, the rate of increase in aconitase activity in a 2-ketoglutarate dehydrogenase (citK) mutant and in a citrate synthase (citA) mutant was significantly less than in an isogenic wild-type strain. This is apparently due to a failure to deplete 2-ketoglutarate and accumulate citrate. These metabolites might act as negative and positive effectors of citB expression, respectively. Mutations known to block sporulation at an early stage (spo0H and spo0B) had no appreciable effect on citB expression or aconitase activity. These results suggest that appearance of aconitase is stimulated by conditions that induce sporulation but is independent of certain gene products thought to act at an early stage of sporulation.
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Selected References
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- Amarasingham C. R., Davis B. D. Regulation of alpha-ketoglutarate dehydrogenase formation in Escherichia coli. J Biol Chem. 1965 Sep;240(9):3664–3668. [PubMed] [Google Scholar]
- Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Dingman D. W., Sonenshein A. L. Purification of aconitase from Bacillus subtilis and correlation of its N-terminal amino acid sequence with the sequence of the citB gene. J Bacteriol. 1987 Jul;169(7):3062–3067. doi: 10.1128/jb.169.7.3062-3067.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donnelly C. E., Sonenshein A. L. Promoter-probe plasmid for Bacillus subtilis. J Bacteriol. 1984 Mar;157(3):965–967. doi: 10.1128/jb.157.3.965-967.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubnau D., Davidoff-Abelson R. Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol. 1971 Mar 14;56(2):209–221. doi: 10.1016/0022-2836(71)90460-8. [DOI] [PubMed] [Google Scholar]
- Ferrari E., Henner D. J., Hoch J. A. Isolation of Bacillus subtilis genes from a charon 4A library. J Bacteriol. 1981 Apr;146(1):430–432. doi: 10.1128/jb.146.1.430-432.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fisher S. H., Magasanik B. 2-Ketoglutarate and the regulation of aconitase and histidase formation in Bacillus subtilis. J Bacteriol. 1984 Apr;158(1):379–382. doi: 10.1128/jb.158.1.379-382.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fisher S. H., Sonenshein A. L. Glutamine-requiring mutants of Bacillus subtilis. Biochem Biophys Res Commun. 1977 Dec 7;79(3):987–995. doi: 10.1016/0006-291x(77)91207-4. [DOI] [PubMed] [Google Scholar]
- Fortnagel P., Freese E. Analysis of sporulation mutants. II. Mutants blocked in the citric acid cycle. J Bacteriol. 1968 Apr;95(4):1431–1438. doi: 10.1128/jb.95.4.1431-1438.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fortnagel P. The regulation of aconitase and isocitrate dehydrogenase in sporulation mutants of Bacillus subtilis. Biochim Biophys Acta. 1970 Nov 24;222(2):290–298. doi: 10.1016/0304-4165(70)90116-9. [DOI] [PubMed] [Google Scholar]
- Freese E. B., Vasantha N., Freese E. Induction of sporulation in developmental mutants of Bacillus subtilis. Mol Gen Genet. 1979 Feb 16;170(1):67–74. doi: 10.1007/BF00268581. [DOI] [PubMed] [Google Scholar]
- Haldenwang W. G., Losick R. Novel RNA polymerase sigma factor from Bacillus subtilis. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7000–7004. doi: 10.1073/pnas.77.12.7000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanson R. S., Cox D. P. Effect of different nutritional conditions on the synthesis of tricarboxylic acid cycle enzymes. J Bacteriol. 1967 Jun;93(6):1777–1787. doi: 10.1128/jb.93.6.1777-1787.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heineken F. G., O'Connor R. J. Continuous culture studies on the biosynthesis of alkaline protease, neutral protease and -amylase by Bacillus subtilis NRRL-B3411. J Gen Microbiol. 1972 Nov;73(1):35–44. doi: 10.1099/00221287-73-1-35. [DOI] [PubMed] [Google Scholar]
- Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
- Igo M. M., Losick R. Regulation of a promoter that is utilized by minor forms of RNA polymerase holoenzyme in Bacillus subtilis. J Mol Biol. 1986 Oct 20;191(4):615–624. doi: 10.1016/0022-2836(86)90449-3. [DOI] [PubMed] [Google Scholar]
- Johnson W. C., Moran C. P., Jr, Losick R. Two RNA polymerase sigma factors from Bacillus subtilis discriminate between overlapping promoters for a developmentally regulated gene. Nature. 1983 Apr 28;302(5911):800–804. doi: 10.1038/302800a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Mitani T., Heinze J. E., Freese E. Induction of sporulation in Bacillus subtilis by decoyinine or hadacidin. Biochem Biophys Res Commun. 1977 Aug 8;77(3):1118–1125. doi: 10.1016/s0006-291x(77)80094-6. [DOI] [PubMed] [Google Scholar]
- Nicholson W. L., Chambliss G. H. Isolation and characterization of a cis-acting mutation conferring catabolite repression resistance to alpha-amylase synthesis in Bacillus subtilis. J Bacteriol. 1985 Mar;161(3):875–881. doi: 10.1128/jb.161.3.875-881.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohné M. Regulation of aconitase synthesis in Bacillus subtilis: induction, feedback repression, and catabolite repression. J Bacteriol. 1974 Mar;117(3):1295–1305. doi: 10.1128/jb.117.3.1295-1305.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ollington J. F., Haldenwang W. G., Huynh T. V., Losick R. Developmentally regulated transcription in a cloned segment of the Bacillus subtilis chromosome. J Bacteriol. 1981 Aug;147(2):432–442. doi: 10.1128/jb.147.2.432-442.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenkrantz M. S., Dingman D. W., Sonenshein A. L. Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine. J Bacteriol. 1985 Oct;164(1):155–164. doi: 10.1128/jb.164.1.155-164.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schreier H. J., Sonenshein A. L. Altered regulation of the glnA gene in glutamine synthetase mutants of Bacillus subtilis. J Bacteriol. 1986 Jul;167(1):35–43. doi: 10.1128/jb.167.1.35-43.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sonenshein A. L., Cami B., Brevet J., Cote R. Isolation and characterization of rifampin-resistant and streptolydigin-resistant mutants of Bacillus subtilis with altered sporulation properties. J Bacteriol. 1974 Oct;120(1):253–265. doi: 10.1128/jb.120.1.253-265.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uratani-Wong B., Lopez J. M., Freese E. Induction of citric acid cycle enzymes during initiation of sporulation by guanine nucleotide deprivation. J Bacteriol. 1981 Apr;146(1):337–344. doi: 10.1128/jb.146.1.337-344.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamazaki H., Ohmura K., Nakayama A., Takeichi Y., Otozai K., Yamasaki M., Tamura G., Yamane K. Alpha-amylase genes (amyR2 and amyE+) from an alpha-amylase-hyperproducing Bacillus subtilis strain: molecular cloning and nucleotide sequences. J Bacteriol. 1983 Oct;156(1):327–337. doi: 10.1128/jb.156.1.327-337.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Zuber P., Losick R. Use of a lacZ fusion to study the role of the spoO genes of Bacillus subtilis in developmental regulation. Cell. 1983 Nov;35(1):275–283. doi: 10.1016/0092-8674(83)90230-1. [DOI] [PubMed] [Google Scholar]