Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Oct 1;17(19):5776–5782. doi: 10.1093/emboj/17.19.5776

sigmaR, an RNA polymerase sigma factor that modulates expression of the thioredoxin system in response to oxidative stress in Streptomyces coelicolor A3(2).

M S Paget 1, J G Kang 1, J H Roe 1, M J Buttner 1
PMCID: PMC1170905  PMID: 9755177

Abstract

We have identified an RNA polymerase sigma factor, sigmaR, that is part of a system that senses and responds to thiol oxidation in the Gram-positive, antibiotic-producing bacterium Streptomyces coelicolor A3(2). Deletion of the gene (sigR) encoding sigmaR caused sensitivity to the thiol-specific oxidant diamide and to the redox cycling compounds menadione and plumbagin. This correlated with reduced levels of disulfide reductase activity and an inability to induce this activity on exposure to diamide. The trxBA operon, encoding thioredoxin reductase and thioredoxin, was found to be under the direct control of sigmaR. trxBA is transcribed from two promoters, trxBp1 and trxBp2, separated by 5-6 bp. trxBp1 is transiently induced at least 50-fold in response to diamide treatment in a sigR-dependent manner. Purified sigmaR directed transcription from trxBp1 in vitro, indicating that trxBp1 is a target for sigmaR. Transcription of sigR itself initiates at two promoters, sigRp1 and sigRp2, which are separated by 173 bp. The sigRp2 transcript was undetectable in a sigR-null mutant, and purified sigmaR could direct transcription from sigRp2 in vitro, indicating that sigR is positively autoregulated. Transcription from sigRp2 was also transiently induced (70-fold) following treatment with diamide. We propose a model in which sigmaR induces expression of the thioredoxin system in response to cytoplasmic disulfide bond formation. Upon reestablishment of normal thiol levels, sigmaR activity is switched off, resulting in down-regulation of trxBA and sigR. We present evidence that the sigmaR system also functions in the actinomycete pathogen Mycobacterium tuberculosis.

Full Text

The Full Text of this article is available as a PDF (318.3 KB).

Selected References

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

  1. Aharonowitz Y., Av-Gay Y., Schreiber R., Cohen G. Characterization of a broad-range disulfide reductase from Streptomyces clavuligerus and its possible role in beta-lactam antibiotic biosynthesis. J Bacteriol. 1993 Feb;175(3):623–629. doi: 10.1128/jb.175.3.623-629.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown K. L., Wood S., Buttner M. J. Isolation and characterization of the major vegetative RNA polymerase of Streptomyces coelicolor A3(2); renaturation of a sigma subunit using GroEL. Mol Microbiol. 1992 May;6(9):1133–1139. doi: 10.1111/j.1365-2958.1992.tb01551.x. [DOI] [PubMed] [Google Scholar]
  3. Buttner M. J., Chater K. F., Bibb M. J. Cloning, disruption, and transcriptional analysis of three RNA polymerase sigma factor genes of Streptomyces coelicolor A3(2). J Bacteriol. 1990 Jun;172(6):3367–3378. doi: 10.1128/jb.172.6.3367-3378.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buttner M. J., Smith A. M., Bibb M. J. At least three different RNA polymerase holoenzymes direct transcription of the agarase gene (dagA) of Streptomyces coelicolor A3(2). Cell. 1988 Feb 26;52(4):599–607. doi: 10.1016/0092-8674(88)90472-2. [DOI] [PubMed] [Google Scholar]
  5. Chakraburtty R., Bibb M. The ppGpp synthetase gene (relA) of Streptomyces coelicolor A3(2) plays a conditional role in antibiotic production and morphological differentiation. J Bacteriol. 1997 Sep;179(18):5854–5861. doi: 10.1128/jb.179.18.5854-5861.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christman M. F., Morgan R. W., Jacobson F. S., Ames B. N. Positive control of a regulon for defenses against oxidative stress and some heat-shock proteins in Salmonella typhimurium. Cell. 1985 Jul;41(3):753–762. doi: 10.1016/s0092-8674(85)80056-8. [DOI] [PubMed] [Google Scholar]
  7. Cohen G., Yanko M., Mislovati M., Argaman A., Schreiber R., Av-Gay Y., Aharonowitz Y. Thioredoxin-thioredoxin reductase system of Streptomyces clavuligerus: sequences, expression, and organization of the genes. J Bacteriol. 1993 Aug;175(16):5159–5167. doi: 10.1128/jb.175.16.5159-5167.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cole S. T., Brosch R., Parkhill J., Garnier T., Churcher C., Harris D., Gordon S. V., Eiglmeier K., Gas S., Barry C. E., 3rd Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998 Jun 11;393(6685):537–544. doi: 10.1038/31159. [DOI] [PubMed] [Google Scholar]
  9. Derman A. I., Prinz W. A., Belin D., Beckwith J. Mutations that allow disulfide bond formation in the cytoplasm of Escherichia coli. Science. 1993 Dec 10;262(5140):1744–1747. doi: 10.1126/science.8259521. [DOI] [PubMed] [Google Scholar]
  10. Hassan H. M., Fridovich I. Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. Arch Biochem Biophys. 1979 Sep;196(2):385–395. doi: 10.1016/0003-9861(79)90289-3. [DOI] [PubMed] [Google Scholar]
  11. Hillemann D., Pühler A., Wohlleben W. Gene disruption and gene replacement in Streptomyces via single stranded DNA transformation of integration vectors. Nucleic Acids Res. 1991 Feb 25;19(4):727–731. doi: 10.1093/nar/19.4.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hindle Z., Smith C. P. Substrate induction and catabolite repression of the Streptomyces coelicolor glycerol operon are mediated through the GylR protein. Mol Microbiol. 1994 Jun;12(5):737–745. doi: 10.1111/j.1365-2958.1994.tb01061.x. [DOI] [PubMed] [Google Scholar]
  13. Holmgren A. Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and thymus and studies of its function in disulfide reduction. J Biol Chem. 1977 Jul 10;252(13):4600–4606. [PubMed] [Google Scholar]
  14. Holmgren A. Reduction of disulfides by thioredoxin. Exceptional reactivity of insulin and suggested functions of thioredoxin in mechanism of hormone action. J Biol Chem. 1979 Sep 25;254(18):9113–9119. [PubMed] [Google Scholar]
  15. Hwang C., Sinskey A. J., Lodish H. F. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992 Sep 11;257(5076):1496–1502. doi: 10.1126/science.1523409. [DOI] [PubMed] [Google Scholar]
  16. Jones G. H., Paget M. S., Chamberlin L., Buttner M. J. Sigma-E is required for the production of the antibiotic actinomycin in Streptomyces antibioticus. Mol Microbiol. 1997 Jan;23(1):169–178. doi: 10.1046/j.1365-2958.1997.2001566.x. [DOI] [PubMed] [Google Scholar]
  17. Kang J. G., Hahn M. Y., Ishihama A., Roe J. H. Identification of sigma factors for growth phase-related promoter selectivity of RNA polymerases from Streptomyces coelicolor A3(2). Nucleic Acids Res. 1997 Jul 1;25(13):2566–2573. doi: 10.1093/nar/25.13.2566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim F. J., Kim H. P., Hah Y. C., Roe J. H. Differential expression of superoxide dismutases containing Ni and Fe/Zn in Streptomyces coelicolor. Eur J Biochem. 1996 Oct 1;241(1):178–185. doi: 10.1111/j.1432-1033.1996.0178t.x. [DOI] [PubMed] [Google Scholar]
  19. Kosower N. S., Kosower E. M. Diamide: an oxidant probe for thiols. Methods Enzymol. 1995;251:123–133. doi: 10.1016/0076-6879(95)51116-4. [DOI] [PubMed] [Google Scholar]
  20. Liochev S. I., Hausladen A., Beyer W. F., Jr, Fridovich I. NADPH: ferredoxin oxidoreductase acts as a paraquat diaphorase and is a member of the soxRS regulon. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1328–1331. doi: 10.1073/pnas.91.4.1328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lonetto M. A., Brown K. L., Rudd K. E., Buttner M. J. Analysis of the Streptomyces coelicolor sigE gene reveals the existence of a subfamily of eubacterial RNA polymerase sigma factors involved in the regulation of extracytoplasmic functions. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7573–7577. doi: 10.1073/pnas.91.16.7573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MacNeil D. J., Occi J. L., Gewain K. M., MacNeil T., Gibbons P. H., Ruby C. L., Danis S. J. Complex organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase. Gene. 1992 Jun 15;115(1-2):119–125. doi: 10.1016/0378-1119(92)90549-5. [DOI] [PubMed] [Google Scholar]
  23. Miranda-Vizuete A., Rodríguez-Ariza A., Toribio F., Holmgren A., López-Barea J., Pueyo C. The levels of ribonucleotide reductase, thioredoxin, glutaredoxin 1, and GSH are balanced in Escherichia coli K12. J Biol Chem. 1996 Aug 9;271(32):19099–19103. doi: 10.1074/jbc.271.32.19099. [DOI] [PubMed] [Google Scholar]
  24. Muller E. G. A glutathione reductase mutant of yeast accumulates high levels of oxidized glutathione and requires thioredoxin for growth. Mol Biol Cell. 1996 Nov;7(11):1805–1813. doi: 10.1091/mbc.7.11.1805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Newton G. L., Arnold K., Price M. S., Sherrill C., Delcardayre S. B., Aharonowitz Y., Cohen G., Davies J., Fahey R. C., Davis C. Distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes. J Bacteriol. 1996 Apr;178(7):1990–1995. doi: 10.1128/jb.178.7.1990-1995.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Paget M. S., Hintermann G., Smith C. P. Construction and application of streptomycete promoter probe vectors which employ the Streptomyces glaucescens tyrosinase-encoding gene as reporter. Gene. 1994 Aug 19;146(1):105–110. doi: 10.1016/0378-1119(94)90842-7. [DOI] [PubMed] [Google Scholar]
  27. Prinz W. A., Aslund F., Holmgren A., Beckwith J. The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm. J Biol Chem. 1997 Jun 20;272(25):15661–15667. doi: 10.1074/jbc.272.25.15661. [DOI] [PubMed] [Google Scholar]
  28. Redenbach M., Kieser H. M., Denapaite D., Eichner A., Cullum J., Kinashi H., Hopwood D. A. A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol. 1996 Jul;21(1):77–96. doi: 10.1046/j.1365-2958.1996.6191336.x. [DOI] [PubMed] [Google Scholar]
  29. Scharf C., Riethdorf S., Ernst H., Engelmann S., Völker U., Hecker M. Thioredoxin is an essential protein induced by multiple stresses in Bacillus subtilis. J Bacteriol. 1998 Apr;180(7):1869–1877. doi: 10.1128/jb.180.7.1869-1877.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Storz G., Tartaglia L. A., Ames B. N. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. Science. 1990 Apr 13;248(4952):189–194. doi: 10.1126/science.2183352. [DOI] [PubMed] [Google Scholar]
  31. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  32. Tao K. oxyR-dependent induction of Escherichia coli grx gene expression by peroxide stress. J Bacteriol. 1997 Sep;179(18):5967–5970. doi: 10.1128/jb.179.18.5967-5970.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wieles B., Ottenhoff T. H., Steenwijk T. M., Franken K. L., de Vries R. R., Langermans J. A. Increased intracellular survival of Mycobacterium smegmatis containing the Mycobacterium leprae thioredoxin-thioredoxin reductase gene. Infect Immun. 1997 Jul;65(7):2537–2541. doi: 10.1128/iai.65.7.2537-2541.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zheng M., Aslund F., Storz G. Activation of the OxyR transcription factor by reversible disulfide bond formation. Science. 1998 Mar 13;279(5357):1718–1721. doi: 10.1126/science.279.5357.1718. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES