Skip to main content
SpringerLink
Log in

Characterization of the alternative sigma factor σG inStreptomyces coelicolor A3(2)

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Using the previously established two-plasmid system for the identification of promoters recognized by a particular sigma factor, we identified two positive DNA fragments that were active only after inducedsigG, encoding σ factor σG ofStreptomyces coelicolor A3(2). High-resolution S1-nuclease mapping in theEscherichia coli two-plasmid system identified potential promoters, PG45 and PG54, whose sequences were similar to the consensus sequence ofBacillus subtilis promoters recognized by the general stress-response σ factor σB. However, both putative σG-dependent promoters were not active inS. coelicolor. Sequence analysis of the regions potentially governed by the promoters revealed a gene encoding a hypothetical protein SCO5555 and therrnE gene encoding rRNA operon. To confirm thatsigG encodes σ factor, the σG protein was overproduced inE. coli and purified. In anin vitro transcription assay, σG, after complementation withS. coelicolor core RNA polymerase, was able to recognize both σG-dependent promoters and initiate transcription.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ausubel F.M., Brent R., Kingston R.E., Moore D.O., Seidman J.S., Smith J.A., Struhl K.:Current Protocols in Molecular Biology. Wiley, New York 1995.

    Google Scholar 

  • Baylis H.A., Bibb M.J.: Organization of the ribosomal RNA genes inStreptomyces coelicolor A3(2).Mol.Gen.Genet. 211, 191–196 (1988a).

    Article  PubMed  CAS  Google Scholar 

  • Baylis H.A., Bibb M.J.. Transcriptional analysis of the 16S rRNA gene of therrnD gene set ofStreptomyces coelicolor A3(2).Mol. Microbiol. 2, 569–579 (1988b).

    Article  PubMed  CAS  Google Scholar 

  • Bentley S.D., Chater K.F., Cerdeno-Tarraga A.M., Challis G.L., Thomson N.R., James K.D., Harris D.E., Quail M.A., Kieser H., Harper D., Bateman A., Brown S., Chandra G., Chen C.W., Collins M., Cronin A., Fraser A., Goble A., Hidalgo J., Hornsby T., Howarth S., Huang C.H., Kieser T., Larke L., Murphy L., Oliver K., O’Neil S., Rabbinowitsch E., Rajandream M.A., Rutherford K., Rutter S., Seeger K., Saunders D., Sharp S., Squares R., Squares S., Taylor K., Warren T., Wietzorrek A., Woodward J., Barrell B.G., Parkhill J., Hopwood D.A.: Complete genome sequence of the model actinomyceteStreptomyces coelicolor A3(2).Nature 417, 141–147 (2002).

    Article  PubMed  Google Scholar 

  • Chater K.F.: Developmental decisions during sporulation in the aerial mycelium inStreptomyces, pp. 33–48 in Y.V. Brun, L.J. Shimkets (Eds):Prokaryotic Development. American Society for Microbiology Press, Washington (DC) 2000.

    Google Scholar 

  • Cho Y.-H., Lee E.-J., Ahn B.-E., Roe J.-H.: SigB, an RNA polymerase σ factor required for osmoprotection and proper differentiation ofStreptomyces coelicolor.Mol.Microbiol. 42, 205–214 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Hecker M., Schumann W., Volker U.: Heat-shock and general stress response inBacillus subtilis.Mol.Microbiol. 19, 417–428 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Homerova D., Ševčiková J., Kormanec J.: Characterization of theStreptomyces coelicolor A3(2)wblE gene, encoding a homologue of the sporulation transcription factor.Folia Microbiol. 48, 489–495 (2003).

    Article  CAS  Google Scholar 

  • Ikeda H., Ishikawa J., Hanamoto A., Shinose M., Kikuchi H., Shiba T., Sabaki Y., Hattori Y., Omura S.: Complete genome sequence and comparative analysis of the industrial microorganismStreptomyces avermitilis.Nature Biotechnol. 21, 526–531 (2003).

    Article  Google Scholar 

  • Kato F., Hino T., Nakaja A., Tanaka M., Koyama Y.: Carotenoid synthesis inStreptomyces setonii ISP5395 is induced by the genecrtS, whose product is similar to σ factor.Mol.Gen.Genet. 247, 387–390 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Kelemen G.H., Viollier P.H., Tenor J.L., Marri L., Buttner M.J., Thompson C.J.: A connection between stress and development in the multicellular prokaryoteStreptomyces coelicolor A3(2).Mol.Microbiol. 40, 804–814 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Kieser T., Bibb M.J., Buttner M.J., Chater K.F., Hopwood D.A.:Practical Streptomyces Genetics. The John Innes Foundation, Norwich (UK) 2000.

  • Kormanec J.: Analyzing the developmental expression of σ factors with SI-nuclease mapping, pp. 481–494 in C.H. Chein (Ed.):Nuclease Methods and Protocols. Methods in Molecular Biology 160. Humana Press, Totowa (USA) 2001.

    Chapter  Google Scholar 

  • Kormanec J., Ševčikova B.: Identification and transcriptional analysis of a cold shock-inducible gene,cspA, inStreptomyces coelicolor A3(2).Mol.Gen.Genet. 264, 251–256 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Kormanec J., Ševiková B., Sprušansky O., Benada O., Kofroňova O., Nováková R., Řežuchová B., Potúčková L., Homerova D.: TheStreptomyces aureofaciens homologue of thewhiB gene is essential for sporulation and its expression correlates with the developmental stage.Folia Microbiol. 43, 605–612 (1998).

    Article  CAS  Google Scholar 

  • Kormanec J., Homerova D., Barak I., Ševciková B.: A new gene,sigG, encoding a putative alternative σ factor ofStreptomyces coelicolor A3(2).FEMS Microbiol.Lett. 172, 153–158 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Kormanec J., Ševčiková B., Halgašova N., Knirschová R., Řežuchova B.: Identification and transcriptional characterization of the gene encoding the stress-response σ factor σH inStreptomyces coelicolor A3(2).FEMS Microbiol.Lett. 189, 31–38 (2000).

    PubMed  CAS  Google Scholar 

  • Kormanec J., Homerová D., Ševčiková B., Řežuchová B.: A method for isolation of small DNA fragments from agarose and polyacrylamide gels.Anal.Biochem. 293, 138–139 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Kundu T.K., Kusano S., Ishihama A.: Promoter selectivity ofEscherichia coli RNA polymerase σF holoenzyme involved in transcription of flagellar and chemotaxis genes.J.Bacteriol. 179, 4264–4269 (1997).

    PubMed  CAS  Google Scholar 

  • Lee H.-S., Ohnishi Z., Horinouchi S.: A σB-like factor responsible for caroteonoid biosynthesis inStreptomyces griseus.J.Mol.Microbiol.Biotechnol. 3, 95–101 (2001).

    PubMed  CAS  Google Scholar 

  • Maxam A.M., Gilbert W.: Sequencing end-labeled DNA with base specific chemical cleavages.Meth.Enzymol. 65, 449–560 (1980).

    Article  Google Scholar 

  • Nováková R., Ševčiková B., Kormanec J.: A method for the identification of promoters recognized by RNA polymerase containing a particular σ factor: cloning of a developmentally regulated promoter and corresponding gene directed by theStreptomyces aureofaciens σ factor RpoZ.Gene 208, 43–50 (1998).

    Article  PubMed  Google Scholar 

  • Park S.F., Stirling D.A., Hulton C.S.J., Booth I.R., Higgins C.F., Stewart G.S.A.B.: A novel, noninvasive promoter probe vector: cloning of the osmoregulatedproU promoter ofEscherichia coli K12.Mol.Microbiol. 3, 1011–1023 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Potučkova L., Kelemen G.H., Findlay K.C., Lonetto M.A., Buttner M.J., Kormanec J.: A new RNA polymerase σ factor, σF, is required for the late stages of morphological differentiation inStreptomyces spp.Mol.Microbiol. 17, 37–48 (1995).

    Article  PubMed  Google Scholar 

  • Price C.W.: General stress response, pp. 369–384 in A.L. Sonenshein (Ed.):Bacillus subtilis and Its Closest Releatives: from Genes to Cells. American Society for Microbiology, Washington (DC) 2002.

    Google Scholar 

  • Ševčikova B., Benada O., Kofroňová O., Kormanec J.: Stress-response σ factor σH is essential for morphological differentiation ofStreptomyces coelicolor A3(2).Arch.Microbiol. 177, 98–106 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Ševčíková B., Kormanec J.: Activity of theStreptomyces coelicolor stress-response σ factor σH is regulated by an anti-σ factor.FEMS Microbiol.Lett. 209, 229–235 (2002).

    PubMed  Google Scholar 

  • Viollier P.H., Kelemen G.H., Dale G.E., Nguyen K.T., Buttner M.J., Thompson C.J.: Specialized osmotic stress response systems involve multiple SigB-like σ factors inStreptomyces coelicolor.Mol.Microbiol. 47, 699–714 (2003a).

    Article  PubMed  CAS  Google Scholar 

  • Viollier P.H., Weihofen A., Folcher M., Thompson C.J.: Post-transcriptional regulation of theStreptomyces coelicolor stress responsive σ factor, SigH, involves translational control, proteolytic processing, and an anti-σ factor homologue.J.Mol.Biol. 325, 637–649 (2003b).

    Article  PubMed  CAS  Google Scholar 

  • van Wezel G.P., Krab I.M., Douthwaite S., Bibb M.J., Vijgenboom E., Bosch L.: Transcriptional analysis of theStreptomyces coelicolor A3(2)rrnA operon.Microbiology SGM 140, 3357–3365 (1994).

    Google Scholar 

  • Wright F., Bibb M.J.: Codon usage in the G+C richStreptomyces genome.Gene 113, 55–65 (1992).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, 845 51, Bratislava, Slovakia

    B. Ševčíková, V. Mazuráková & J. Kormanec

Authors
  1. B. Ševčíková
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. V. Mazuráková
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Kormanec
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by grant 2/3010/23 from theSlovak Academy of Sciences.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ševčíková, B., Mazuráková, V. & Kormanec, J. Characterization of the alternative sigma factor σG inStreptomyces coelicolor A3(2). Folia Microbiol 50, 47–58 (2005). https://doi.org/10.1007/BF02931293

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02931293

Keywords

Search

Navigation