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. 1985 Oct;164(1):155–164. doi: 10.1128/jb.164.1.155-164.1985

Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine.

M S Rosenkrantz, D W Dingman, A L Sonenshein
PMCID: PMC214224  PMID: 2413006

Abstract

The activity of aconitase in Bacillus subtilis is greatly reduced in cells cultured in media containing rapidly metabolized carbon sources (e.g., glucose). Thus, expression of this enzyme appears to be subject to a form of catabolite repression. Since the product of the citB gene of B. subtilis is required for aconitase activity, we cloned the wild-type allele of this gene and used this DNA as a probe for transcription of citB in cells grown in various media. The steady-state level of RNA that hybridized to this probe was about 10-fold higher in B. subtilis cells grown in citrate-glutamine medium than in cells grown in glucose-glutamine medium. This result correlates well with the steady-state levels of aconitase activity. Two transcripts were shown to initiate within the cloned DNA; the steady-state level of one of these transcripts varied in the same way as did aconitase activity when cells were grown in media containing different carbon sources. This is the first demonstration of regulation by the carbon source of the level of a vegatative-cell transcript in B. subtilis.

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  1. 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]
  2. Bernlohr R. W., Haddox M. K., Goldberg N. D. Cyclic guanosine 3':5'-monophosphate in Escherichia coli and Bacillus lichenformis. J Biol Chem. 1974 Jul 10;249(13):4329–4331. [PubMed] [Google Scholar]
  3. 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]
  4. Bolivar F. Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules. Gene. 1978 Oct;4(2):121–136. doi: 10.1016/0378-1119(78)90025-2. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Coote J. G. Comparative studies on induction of sporulation and synthesis of inducible enzymes in Bacillus subtilis. J Bacteriol. 1974 Dec;120(3):1102–1108. doi: 10.1128/jb.120.3.1102-1108.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. 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]
  10. Fisher S. H., Magasanik B. Isolation of Bacillus subtilis mutants pleiotropically insensitive to glucose catabolite repression. J Bacteriol. 1984 Mar;157(3):942–944. doi: 10.1128/jb.157.3.942-944.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fisher S. H., Magasanik B. Synthesis of oxaloacetate in Bacillus subtilis mutants lacking the 2-ketoglutarate dehydrogenase enzymatic complex. J Bacteriol. 1984 Apr;158(1):55–62. doi: 10.1128/jb.158.1.55-62.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fisher S. H., Rosenkrantz M. S., Sonenshein A. L. Glutamine synthetase gene of Bacillus subtilis. Gene. 1984 Dec;32(3):427–438. doi: 10.1016/0378-1119(84)90018-0. [DOI] [PubMed] [Google Scholar]
  13. Fisher S. H., Sonenshein A. L. Bacillus subtilis glutamine synthetase mutants pleiotropically altered in glucose catabolite repression. J Bacteriol. 1984 Feb;157(2):612–621. doi: 10.1128/jb.157.2.612-621.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flechtner V. R., Hanson R. S. Coarse and fine control of citrate synthase from Bacillus subtilis. Biochim Biophys Acta. 1969 Jul 30;184(2):252–262. doi: 10.1016/0304-4165(69)90027-0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gilman M. Z., Chamberlin M. J. Developmental and genetic regulation of Bacillus subtilis genes transcribed by sigma 28-RNA polymerase. Cell. 1983 Nov;35(1):285–293. doi: 10.1016/0092-8674(83)90231-3. [DOI] [PubMed] [Google Scholar]
  19. Gitt M. A., Wang L. F., Doi R. H. A strong sequence homology exists between the major RNA polymerase sigma factors of Bacillus subtilis and Escherichia coli. J Biol Chem. 1985 Jun 25;260(12):7178–7185. [PubMed] [Google Scholar]
  20. HANSON H. M., WITOSLAWSKI J. J., CAMPBELL E. H. REVERSIBLE DISRUPTION OF A WAVELENGTH DISCRIMINATION IN PIGEONS FOLLOWING ADMINISTRATION OF PHENIPRAZINE. Toxicol Appl Pharmacol. 1964 Nov;6:690–695. doi: 10.1016/0041-008x(64)90119-x. [DOI] [PubMed] [Google Scholar]
  21. Haldenwang W. G., Lang N., Losick R. A sporulation-induced sigma-like regulatory protein from B. subtilis. Cell. 1981 Feb;23(2):615–624. doi: 10.1016/0092-8674(81)90157-4. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. 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]
  25. Le Grice S. F., Sonenshein A. L. Interaction of Bacillus subtilis RNA polymerase with a chromosomal promoter. J Mol Biol. 1982 Dec 15;162(3):551–564. doi: 10.1016/0022-2836(82)90388-6. [DOI] [PubMed] [Google Scholar]
  26. Lopez J. M., Thoms B. Role of sugar uptake and metabolic intermediates on catabolite repression in Bacillus subtilis. J Bacteriol. 1977 Jan;129(1):217–224. doi: 10.1128/jb.129.1.217-224.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Losick R., Pero J. Cascades of Sigma factors. Cell. 1981 Sep;25(3):582–584. doi: 10.1016/0092-8674(81)90164-1. [DOI] [PubMed] [Google Scholar]
  28. Losick R., Sonenshein A. L. Change in the template specificity of RNA polymerase during sporulation of Bacillus subtilis. Nature. 1969 Oct 4;224(5214):35–37. doi: 10.1038/224035a0. [DOI] [PubMed] [Google Scholar]
  29. Marusyk R., Sergeant A. A simple method for dialysis of small-volume samples. Anal Biochem. 1980 Jul 1;105(2):403–404. doi: 10.1016/0003-2697(80)90477-7. [DOI] [PubMed] [Google Scholar]
  30. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  31. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  32. Morrison D. A. Transformation in Escherichia coli: cryogenic preservation of competent cells. J Bacteriol. 1977 Oct;132(1):349–351. doi: 10.1128/jb.132.1.349-351.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. 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]
  35. Ohné M. Regulation of the dicarboxylic acid part of the citric acid cycle in Bacillus subtilis. J Bacteriol. 1975 Apr;122(1):224–234. doi: 10.1128/jb.122.1.224-234.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Ollington J. F., Losick R. A cloned gene that is turned on at an intermediate stage of spore formation in Bacillus subtilis. J Bacteriol. 1981 Aug;147(2):443–451. doi: 10.1128/jb.147.2.443-451.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Prentki P., Karch F., Iida S., Meyer J. The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene. 1981 Sep;14(4):289–299. doi: 10.1016/0378-1119(81)90161-x. [DOI] [PubMed] [Google Scholar]
  39. Rutberg B., Hoch J. A. Citric acid cycle: gene-enzyme relationships in Bacillus subtilis. J Bacteriol. 1970 Nov;104(2):826–833. doi: 10.1128/jb.104.2.826-833.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scholze H. Studies on aconitase species from Saccharomyces cerevisiae, porcine and bovine heart, obtained by a modified isolation method. Biochim Biophys Acta. 1983 Aug 16;746(3):133–137. doi: 10.1016/0167-4838(83)90066-3. [DOI] [PubMed] [Google Scholar]
  41. Setlow P. Inability of detect cyclic AMP in vegetative or sporulating cells or dormant spores of Bacillus megaterium. Biochem Biophys Res Commun. 1973 May 15;52(2):365–372. doi: 10.1016/0006-291x(73)90720-1. [DOI] [PubMed] [Google Scholar]
  42. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tjian R., Losick R. An immunological assay for the sigma subunit of RNA polymerase in extracts of vegetative and sporulating Bacillus subtilis. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2872–2876. doi: 10.1073/pnas.71.7.2872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wiggs J. L., Gilman M. Z., Chamberlin M. J. Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional sigma factor in vegetative cells. Proc Natl Acad Sci U S A. 1981 May;78(5):2762–2766. doi: 10.1073/pnas.78.5.2762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]

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