Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Dec;178(23):7020–7023. doi: 10.1128/jb.178.23.7020-7023.1996

The yeast two-hybrid system detects interactions between Bacillus subtilis sigmaB regulators.

U Voelker 1, A Voelker 1, W G Haldenwang 1
PMCID: PMC178610  PMID: 8955331

Abstract

SigmaB, the general stress response sigma factor of Bacillus subtilis, is regulated by the products of seven genes (rsbR, S, T, U, V, W, and X) with which it is cotranscribed. Biochemical techniques previously revealed physical associations among RsbW, RsbV, and sigmaB but failed to detect interactions of RsbR, S, T, U, or X with each other or RsbV, RsbW, or sigmaB. Using the yeast two-hybrid system, we have now obtained evidence for such interactions. The yeast reporter system was activated when RsbS was paired with either RsbR or RsbT, RsbR was paired with RsbT, and RsbV was paired with either RsbU or RsbW. In addition, RsbW2 and RsbR2 dimer formation was detected. RsbX failed to show interactions with itself or any of the other sigB operon products.

Full Text

The Full Text of this article is available as a PDF (180.7 KB).

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. Arigoni F., Duncan L., Alper S., Losick R., Stragier P. SpoIIE governs the phosphorylation state of a protein regulating transcription factor sigma F during sporulation in Bacillus subtilis. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3238–3242. doi: 10.1073/pnas.93.8.3238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benson A. K., Haldenwang W. G. Bacillus subtilis sigma B is regulated by a binding protein (RsbW) that blocks its association with core RNA polymerase. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2330–2334. doi: 10.1073/pnas.90.6.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Benson A. K., Haldenwang W. G. Regulation of sigma B levels and activity in Bacillus subtilis. J Bacteriol. 1993 Apr;175(8):2347–2356. doi: 10.1128/jb.175.8.2347-2356.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benson A. K., Haldenwang W. G. The sigma B-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock. J Bacteriol. 1993 Apr;175(7):1929–1935. doi: 10.1128/jb.175.7.1929-1935.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Binnie C., Lampe M., Losick R. Gene encoding the sigma 37 species of RNA polymerase sigma factor from Bacillus subtilis. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5943–5947. doi: 10.1073/pnas.83.16.5943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Dufour A., Haldenwang W. G. Interactions between a Bacillus subtilis anti-sigma factor (RsbW) and its antagonist (RsbV). J Bacteriol. 1994 Apr;176(7):1813–1820. doi: 10.1128/jb.176.7.1813-1820.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dufour A., Voelker U., Voelker A., Haldenwang W. G. Relative levels and fractionation properties of Bacillus subtilis σ(B) and its regulators during balanced growth and stress. J Bacteriol. 1996 Jul;178(13):3701–sigma. doi: 10.1128/jb.178.13.3701-9sigma.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Fields S., Sternglanz R. The two-hybrid system: an assay for protein-protein interactions. Trends Genet. 1994 Aug;10(8):286–292. doi: 10.1016/0168-9525(90)90012-u. [DOI] [PubMed] [Google Scholar]
  13. Guarente L. Strategies for the identification of interacting proteins. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1639–1641. doi: 10.1073/pnas.90.5.1639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kalman S., Duncan M. L., Thomas S. M., Price C. W. Similar organization of the sigB and spoIIA operons encoding alternate sigma factors of Bacillus subtilis RNA polymerase. J Bacteriol. 1990 Oct;172(10):5575–5585. doi: 10.1128/jb.172.10.5575-5585.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kang C. M., Brody M. S., Akbar S., Yang X., Price C. W. Homologous pairs of regulatory proteins control activity of Bacillus subtilis transcription factor sigma(b) in response to environmental stress. J Bacteriol. 1996 Jul;178(13):3846–3853. doi: 10.1128/jb.178.13.3846-3853.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Magnin T., Lord M., Errington J., Yudkin M. D. Establishing differential gene expression in sporulating Bacillus subtilis: phosphorylation of SpoIIAA (anti-anti-sigmaF) alters its conformation and prevents formation of a SpoIIAA/SpoIIAB/ADP complex. Mol Microbiol. 1996 Feb;19(4):901–907. doi: 10.1046/j.1365-2958.1996.434964.x. [DOI] [PubMed] [Google Scholar]
  18. Voelker U., Dufour A., Haldenwang W. G. The Bacillus subtilis rsbU gene product is necessary for RsbX-dependent regulation of sigma B. J Bacteriol. 1995 Jan;177(1):114–122. doi: 10.1128/jb.177.1.114-122.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Voelker U., Voelker A., Haldenwang W. G. Reactivation of the Bacillus subtilis anti-sigma B antagonist, RsbV, by stress- or starvation-induced phosphatase activities. J Bacteriol. 1996 Sep;178(18):5456–5463. doi: 10.1128/jb.178.18.5456-5463.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Wise A. A., Price C. W. Four additional genes in the sigB operon of Bacillus subtilis that control activity of the general stress factor sigma B in response to environmental signals. J Bacteriol. 1995 Jan;177(1):123–133. doi: 10.1128/jb.177.1.123-133.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES