Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Nov;179(21):6778–6787. doi: 10.1128/jb.179.21.6778-6787.1997

Regulation of Bacillus subtilis sigmaH (spo0H) and AbrB in response to changes in external pH.

W M Cosby 1, P Zuber 1
PMCID: PMC179609  PMID: 9352930

Abstract

The RNA polymerase sigma subunit, sigmaH, of Bacillus subtilis is required for the transcription of genes that are induced in late-growth cultures at high cell density, including genes that function in sporulation. The expression of sigmaH-controlled genes is repressed when nutrient broth sporulation medium (Difco sporulation medium [DSM]) is supplemented with high concentrations of glucose and glutamine (DSM-GG), preferred carbon and nitrogen sources of B. subtilis. Under these conditions, the pH of the DSM-GG medium decreases to approximately 5. Raising the pH by the addition of morpholinepropanesulfonic acid (MOPS) or Tris-HCl (pH 7.5) results in a dramatic increase in the expression of lacZ fusions to sigmaH-dependent promoters. Correspondingly, the level of sigmaH protein was higher in cells of late-growth DSM-GG cultures treated with a pH stabilizer. When sigmaH-dependent gene expression was examined in cells bearing a mutation in abrB, encoding the transition state regulator that negatively controls genes transcribed by the sigmaH form of RNA polymerase, derepression was observed as well as an increase in medium pH. Reducing the pH with acetic acid resulted in repression, suggesting that AbrB was not functioning directly in pH-dependent repression but was required to maintain the low medium pH in DSM-GG. AbrB protein levels were high in late-growth, DSM-GG cultures but significantly lower when the pH was raised by Tris-HCl addition. An active tricarboxylic acid (TCA) cycle was required to obtain maximum derepression of sigmaH-dependent transcription, and transcription of the TCA cycle enzyme gene citB was repressed in DSM-GG but derepressed when the pH was artificially raised. The negative effect of low pH on sigmaH-dependent lacZ expression was also observed in unbuffered minimal medium and appeared to be exerted posttranslationally with respect to spo0H expression. However, the addition of amino acids to the medium caused pH-independent repression of both sigmaH-dependent transcription and spo0H-lacZ expression. These results suggest that spo0H transcription or translation is repressed by a mechanism responding to the availability of amino acids whereas spo0H is posttranslationally regulated in response to external pH.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

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

  1. Alper S., Dufour A., Garsin D. A., Duncan L., Losick R. Role of adenosine nucleotides in the regulation of a stress-response transcription factor in Bacillus subtilis. J Mol Biol. 1996 Jul 12;260(2):165–177. doi: 10.1006/jmbi.1996.0390. [DOI] [PubMed] [Google Scholar]
  2. Alper S., Duncan L., Losick R. An adenosine nucleotide switch controlling the activity of a cell type-specific transcription factor in B. subtilis. Cell. 1994 Apr 22;77(2):195–205. doi: 10.1016/0092-8674(94)90312-3. [DOI] [PubMed] [Google Scholar]
  3. Asai K., Kawamura F., Yoshikawa H., Takahashi H. Expression of kinA and accumulation of sigma H at the onset of sporulation in Bacillus subtilis. J Bacteriol. 1995 Nov;177(22):6679–6683. doi: 10.1128/jb.177.22.6679-6683.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumberg S., Harwood C. R. Carbon and nitrogen repression of arginine catabolic enzymes in Bacillus subtilis. J Bacteriol. 1979 Jan;137(1):189–196. doi: 10.1128/jb.137.1.189-196.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benson A. K., Haldenwang W. G. Characterization of a regulatory network that controls sigma B expression in Bacillus subtilis. J Bacteriol. 1992 Feb;174(3):749–757. doi: 10.1128/jb.174.3.749-757.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boylan S. A., Redfield A. R., Brody M. S., Price C. W. Stress-induced activation of the sigma B transcription factor of Bacillus subtilis. J Bacteriol. 1993 Dec;175(24):7931–7937. doi: 10.1128/jb.175.24.7931-7937.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Boylan S. A., Redfield A. R., Price C. W. Transcription factor sigma B of Bacillus subtilis controls a large stationary-phase regulon. J Bacteriol. 1993 Jul;175(13):3957–3963. doi: 10.1128/jb.175.13.3957-3963.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burbulys D., Trach K. A., Hoch J. A. Initiation of sporulation in B. subtilis is controlled by a multicomponent phosphorelay. Cell. 1991 Feb 8;64(3):545–552. doi: 10.1016/0092-8674(91)90238-t. [DOI] [PubMed] [Google Scholar]
  9. Carter H. L., 3rd, Moran C. P., Jr New RNA polymerase sigma factor under spo0 control in Bacillus subtilis. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9438–9442. doi: 10.1073/pnas.83.24.9438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Diederich B., Wilkinson J. F., Magnin T., Najafi M., Erringston J., Yudkin M. D. Role of interactions between SpoIIAA and SpoIIAB in regulating cell-specific transcription factor sigma F of Bacillus subtilis. Genes Dev. 1994 Nov 1;8(21):2653–2663. doi: 10.1101/gad.8.21.2653. [DOI] [PubMed] [Google Scholar]
  11. Dingman D. W., Rosenkrantz M. S., Sonenshein A. L. Relationship between aconitase gene expression and sporulation in Bacillus subtilis. J Bacteriol. 1987 Jul;169(7):3068–3075. doi: 10.1128/jb.169.7.3068-3075.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Dubnau E., Weir J., Nair G., Carter L., 3rd, Moran C., Jr, Smith I. Bacillus sporulation gene spo0H codes for sigma 30 (sigma H). J Bacteriol. 1988 Mar;170(3):1054–1062. doi: 10.1128/jb.170.3.1054-1062.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Duncan L., Alper S., Arigoni F., Losick R., Stragier P. Activation of cell-specific transcription by a serine phosphatase at the site of asymmetric division. Science. 1995 Oct 27;270(5236):641–644. doi: 10.1126/science.270.5236.641. [DOI] [PubMed] [Google Scholar]
  15. Duncan L., Losick R. SpoIIAB is an anti-sigma factor that binds to and inhibits transcription by regulatory protein sigma F from Bacillus subtilis. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2325–2329. doi: 10.1073/pnas.90.6.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fisher S. H., Sonenshein A. L. Control of carbon and nitrogen metabolism in Bacillus subtilis. Annu Rev Microbiol. 1991;45:107–135. doi: 10.1146/annurev.mi.45.100191.000543. [DOI] [PubMed] [Google Scholar]
  17. Fouet A., Jin S. F., Raffel G., Sonenshein A. L. Multiple regulatory sites in the Bacillus subtilis citB promoter region. J Bacteriol. 1990 Sep;172(9):5408–5415. doi: 10.1128/jb.172.9.5408-5415.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fouet A., Sonenshein A. L. A target for carbon source-dependent negative regulation of the citB promoter of Bacillus subtilis. J Bacteriol. 1990 Feb;172(2):835–844. doi: 10.1128/jb.172.2.835-844.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Frisby D., Zuber P. Analysis of the upstream activating sequence and site of carbon and nitrogen source repression in the promoter of an early-induced sporulation gene of Bacillus subtilis. J Bacteriol. 1991 Dec;173(23):7557–7564. doi: 10.1128/jb.173.23.7557-7564.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Frisby D., Zuber P. Mutations in pts cause catabolite-resistant sporulation and altered regulation of spo0H in Bacillus subtilis. J Bacteriol. 1994 May;176(9):2587–2595. doi: 10.1128/jb.176.9.2587-2595.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grossman A. D. Genetic networks controlling the initiation of sporulation and the development of genetic competence in Bacillus subtilis. Annu Rev Genet. 1995;29:477–508. doi: 10.1146/annurev.ge.29.120195.002401. [DOI] [PubMed] [Google Scholar]
  22. Grundy F. J., Henkin T. M. Characterization of the Bacillus subtilis rpsD regulatory target site. J Bacteriol. 1992 Nov;174(21):6763–6770. doi: 10.1128/jb.174.21.6763-6770.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Grundy F. J., Turinsky A. J., Henkin T. M. Catabolite regulation of Bacillus subtilis acetate and acetoin utilization genes by CcpA. J Bacteriol. 1994 Aug;176(15):4527–4533. doi: 10.1128/jb.176.15.4527-4533.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Haldenwang W. G. The sigma factors of Bacillus subtilis. Microbiol Rev. 1995 Mar;59(1):1–30. doi: 10.1128/mr.59.1.1-30.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Healy J., Weir J., Smith I., Losick R. Post-transcriptional control of a sporulation regulatory gene encoding transcription factor sigma H in Bacillus subtilis. Mol Microbiol. 1991 Feb;5(2):477–487. doi: 10.1111/j.1365-2958.1991.tb02131.x. [DOI] [PubMed] [Google Scholar]
  26. Hicks K. A., Grossman A. D. Altering the level and regulation of the major sigma subunit of RNA polymerase affects gene expression and development in Bacillus subtilis. Mol Microbiol. 1996 Apr;20(1):201–212. doi: 10.1111/j.1365-2958.1996.tb02501.x. [DOI] [PubMed] [Google Scholar]
  27. Hofmeister A. E., Londoño-Vallejo A., Harry E., Stragier P., Losick R. Extracellular signal protein triggering the proteolytic activation of a developmental transcription factor in B. subtilis. Cell. 1995 Oct 20;83(2):219–226. doi: 10.1016/0092-8674(95)90163-9. [DOI] [PubMed] [Google Scholar]
  28. Igo M., Lampe M., Ray C., Schafer W., Moran C. P., Jr, Losick R. Genetic studies of a secondary RNA polymerase sigma factor in Bacillus subtilis. J Bacteriol. 1987 Aug;169(8):3464–3469. doi: 10.1128/jb.169.8.3464-3469.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jin S., Sonenshein A. L. Identification of two distinct Bacillus subtilis citrate synthase genes. J Bacteriol. 1994 Aug;176(15):4669–4679. doi: 10.1128/jb.176.15.4669-4679.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jin S., Sonenshein A. L. Transcriptional regulation of Bacillus subtilis citrate synthase genes. J Bacteriol. 1994 Aug;176(15):4680–4690. doi: 10.1128/jb.176.15.4680-4690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Krulwich T. A., Agus R., Schneier M., Guffanti A. A. Buffering capacity of bacilli that grow at different pH ranges. J Bacteriol. 1985 May;162(2):768–772. doi: 10.1128/jb.162.2.768-772.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lewis P. J., Partridge S. R., Errington J. Sigma factors, asymmetry, and the determination of cell fate in Bacillus subtilis. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3849–3853. doi: 10.1073/pnas.91.9.3849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Losick R., Stragier P. Crisscross regulation of cell-type-specific gene expression during development in B. subtilis. Nature. 1992 Feb 13;355(6361):601–604. doi: 10.1038/355601a0. [DOI] [PubMed] [Google Scholar]
  34. Magnuson R., Solomon J., Grossman A. D. Biochemical and genetic characterization of a competence pheromone from B. subtilis. Cell. 1994 Apr 22;77(2):207–216. doi: 10.1016/0092-8674(94)90313-1. [DOI] [PubMed] [Google Scholar]
  35. Marahiel M. A., Zuber P., Czekay G., Losick R. Identification of the promoter for a peptide antibiotic biosynthesis gene from Bacillus brevis and its regulation in Bacillus subtilis. J Bacteriol. 1987 May;169(5):2215–2222. doi: 10.1128/jb.169.5.2215-2222.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nakano M. M., Marahiel M. A., Zuber P. Identification of a genetic locus required for biosynthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis. J Bacteriol. 1988 Dec;170(12):5662–5668. doi: 10.1128/jb.170.12.5662-5668.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Partridge S. R., Errington J. The importance of morphological events and intercellular interactions in the regulation of prespore-specific gene expression during sporulation in Bacillus subtilis. Mol Microbiol. 1993 May;8(5):945–955. doi: 10.1111/j.1365-2958.1993.tb01639.x. [DOI] [PubMed] [Google Scholar]
  38. Perego M., Spiegelman G. B., Hoch J. A. Structure of the gene for the transition state regulator, abrB: regulator synthesis is controlled by the spo0A sporulation gene in Bacillus subtilis. Mol Microbiol. 1988 Nov;2(6):689–699. doi: 10.1111/j.1365-2958.1988.tb00079.x. [DOI] [PubMed] [Google Scholar]
  39. Perkins J. B., Youngman P. J. Construction and properties of Tn917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis. Proc Natl Acad Sci U S A. 1986 Jan;83(1):140–144. doi: 10.1073/pnas.83.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. Solomon J. M., Magnuson R., Srivastava A., Grossman A. D. Convergent sensing pathways mediate response to two extracellular competence factors in Bacillus subtilis. Genes Dev. 1995 Mar 1;9(5):547–558. doi: 10.1101/gad.9.5.547. [DOI] [PubMed] [Google Scholar]
  42. Stragier P., Losick R. Molecular genetics of sporulation in Bacillus subtilis. Annu Rev Genet. 1996;30:297–241. doi: 10.1146/annurev.genet.30.1.297. [DOI] [PubMed] [Google Scholar]
  43. Strauch M. A. Delineation of AbrB-binding sites on the Bacillus subtilis spo0H, kinB, ftsAZ, and pbpE promoters and use of a derived homology to identify a previously unsuspected binding site in the bsuB1 methylase promote. J Bacteriol. 1995 Dec;177(23):6999–7002. doi: 10.1128/jb.177.23.6999-7002.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Strauch M. A., Spiegelman G. B., Perego M., Johnson W. C., Burbulys D., Hoch J. A. The transition state transcription regulator abrB of Bacillus subtilis is a DNA binding protein. EMBO J. 1989 May;8(5):1615–1621. doi: 10.1002/j.1460-2075.1989.tb03546.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Strauch M., Webb V., Spiegelman G., Hoch J. A. The SpoOA protein of Bacillus subtilis is a repressor of the abrB gene. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1801–1805. doi: 10.1073/pnas.87.5.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Voelker U., Voelker A., Maul B., Hecker M., Dufour A., Haldenwang W. G. Separate mechanisms activate sigma B of Bacillus subtilis in response to environmental and metabolic stresses. J Bacteriol. 1995 Jul;177(13):3771–3780. doi: 10.1128/jb.177.13.3771-3780.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Völker U., Engelmann S., Maul B., Riethdorf S., Völker A., Schmid R., Mach H., Hecker M. Analysis of the induction of general stress proteins of Bacillus subtilis. Microbiology. 1994 Apr;140(Pt 4):741–752. doi: 10.1099/00221287-140-4-741. [DOI] [PubMed] [Google Scholar]
  48. Weir J., Predich M., Dubnau E., Nair G., Smith I. Regulation of spo0H, a gene coding for the Bacillus subtilis sigma H factor. J Bacteriol. 1991 Jan;173(2):521–529. doi: 10.1128/jb.173.2.521-529.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yang X., Kang C. M., Brody M. S., Price C. W. Opposing pairs of serine protein kinases and phosphatases transmit signals of environmental stress to activate a bacterial transcription factor. Genes Dev. 1996 Sep 15;10(18):2265–2275. doi: 10.1101/gad.10.18.2265. [DOI] [PubMed] [Google Scholar]
  50. Youngman P., Perkins J. B., Losick R. A novel method for the rapid cloning in Escherichia coli of Bacillus subtilis chromosomal DNA adjacent to Tn917 insertions. Mol Gen Genet. 1984;195(3):424–433. doi: 10.1007/BF00341443. [DOI] [PubMed] [Google Scholar]
  51. Zuber P., Losick R. Role of AbrB in Spo0A- and Spo0B-dependent utilization of a sporulation promoter in Bacillus subtilis. J Bacteriol. 1987 May;169(5):2223–2230. doi: 10.1128/jb.169.5.2223-2230.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES