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Identification of different promoters in the absA1–absA2 two-component system, a negative regulator of antibiotic production in Streptomyces coelicolor

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Abstract

The absA1–absA2 genes encode a two-component system that negatively regulates the transcription of multiple antibiotic gene clusters of Streptomyces coelicolor. The microarray dataset time series of a S. coelicolor M145 bioreactor culture indicated that the transcription values of absA2 were approximately four times higher than those of absA1 throughout the time course of the culture. The co-transcription of absA1 and absA2 genes has been previously shown, although an independent absA2 promoter was not detected. In this study, we show by different technical approaches that the absA1–absA2 operon is transcribed from at least two promoters, the first producing a read-through transcript that includes both absA1 and absA2 genes and the second including only the absA2 gene. An absA2 mRNA 5′ end was mapped by primer extension and confirmed as TSS by deep sequencing in combination with TEX. Promoter-probe analyses detected promoter activity in both the absA1 and absA2 upstream regions. The absA2 upstream region showed a higher promoter activity, at least sevenfold higher than that of absA1. Furthermore, the absA2 gene may contain at least two additional promoters as shown by deep sequencing analyses. All together this work contributes to the understanding of the complex transcriptional regulation of these antibiotic regulators genes in S. coelicolor.

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References

  • Adamidis T, Riggle P, Champness W (1990) Mutations in a new Streptomyces coelicolor locus which globally block antibiotic biosynthesis but not sporulation. J Bacteriol 172:2962–2969

    PubMed  CAS  Google Scholar 

  • Anderson TB, Brian P, Champness WC (2001) Genetic and transcriptional analysis of absA, an antibiotic gene cluster-linked two-component system that regulates multiple antibiotics in Streptomyces coelicolor. Mol Microbiol 39:553–566

    Article  PubMed  CAS  Google Scholar 

  • Batchelor E, Goulian M (2003) Robustness and the cycle of phosphorylation and dephosphorylation in a two-component regulatory system. Proc Natl Acad Sci USA 100:691–696

    Article  PubMed  CAS  Google Scholar 

  • Bibb MJ (2005) Regulation of secondary metabolism in streptomycetes. Curr Opin Microbiol 8:208–215

    Article  PubMed  CAS  Google Scholar 

  • Brian P, Riggle PJ, Santos RA, Champness WC (1996) Global negative regulation of Streptomyces coelicolor antibiotic synthesis mediated by an absA-encoded putative signal transduction system. J Bacteriol 178:3221–3231

    PubMed  CAS  Google Scholar 

  • Champness W, Riggle P, Adamidis T, Vandervere P (1992) Identification of Streptomyces coelicolor genes involved in regulation of antibiotic synthesis. Gene 115:55–60

    Article  PubMed  CAS  Google Scholar 

  • Demain AL (2009) Antibiotics: natural products essential to human health. Med Res Rev 29:821–842

    Article  PubMed  CAS  Google Scholar 

  • Goulian M (2010) Two-component signaling circuit structure and properties. Curr Opin Microbiol 13:184–189

    Article  PubMed  CAS  Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Article  PubMed  CAS  Google Scholar 

  • Hopwood DA (2007) Streptomyces in nature and medicine. Oxford University Press, New York

    Google Scholar 

  • Hutchings MI, Hoskisson PA, Chandra G, Buttner MJ (2004) Sensing and responding to diverse extracellular signals? Analysis of the sensor kinases and response regulators of Streptomyces coelicolor A3(2). Microbiology 150:2795–2806

    Article  PubMed  CAS  Google Scholar 

  • Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. The John Innes Foundation, Norwich

    Google Scholar 

  • Kröger C, Dillon SC, Cameron AD, Papenfort K, Sivasankaran SK, Hokamp K, Chao Y, Sittka A, Hébrard M, Händler K, Colgan A, Leekitcharoenphon P, Langridge GC, Lohan AJ, Loftus B, Lucchini S, Ussery DW, Dorman CJ, Thomson NR, Vogel J, Hinton JC (2012) The transcriptional landscape and small RNAs of Salmonella enterica serovar Typhimurium. Proc Natl Acad Sci USA 109:E1277–E1286

    Article  PubMed  Google Scholar 

  • MacNeil DJ, Gewain KM, Ruby CL, Dezeny G, Gibbons PH, MacNeil T (1992) Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. Gene 111:61–68

    Article  PubMed  CAS  Google Scholar 

  • Martín JF (2004) Phosphate control of the biosynthesis of antibiotics and other secondary metabolites is mediated by the PhoR-PhoP system: an unfinished story. J Bacteriol 186:5197–5201

    Article  PubMed  Google Scholar 

  • Martín JF, Liras P (2010) Engineering of regulatory cascades and networks controlling antibiotic biosynthesis in Streptomyces. Curr Opin Microbiol 13:263–273

    Article  PubMed  Google Scholar 

  • Martín JF, Santos-Beneit F, Rodríguez-García A, Sola-Landa A, Smith MC, Ellingsen TE, Nieselt K, Burroughs NJ, Wellington EM (2012a) Transcriptomic studies of phosphate control of primary and secondary metabolism in Streptomyces coelicolor. Appl Microbiol Biotechnol 95:61–75

    Article  PubMed  Google Scholar 

  • Martín JF, Sola-Landa A, Rodríguez-García A (2012b) Two-component systems in Streptomyces. In: Gross R, Beier D (eds) Two-component systems in bacteria. Caister Academic Press, Würzburg, pp 315–331

    Google Scholar 

  • McKenzie NL, Nodwell JR (2007) Phosphorylated AbsA2 negatively regulates antibiotic production in Streptomyces coelicolor through interactions with pathway-specific regulatory gene promoters. J Bacteriol 189:5284–5292

    Article  PubMed  CAS  Google Scholar 

  • Nicol JW, Helt GA, Blanchard SG Jr, Raja A, Loraine AE (2009) The Integrated Genome Browser: free software for distribution and exploration of genome-scale datasets. Bioinformatics 25:2730–2731

    Article  PubMed  CAS  Google Scholar 

  • Nieselt K, Battke F, Herbig A, Bruheim P, Wentzel A, Jakobsen OM, Sletta H, Alam MT, Merlo ME, Moore J, Omara WA, Morrissey ER, Juarez-Hermosillo MA, Rodríguez-García A, Nentwich M, Thomas L, Iqbal M, Legaie R, Gaze WH, Challis GL, Jansen RC, Dijkhuizen L, Rand DA, Wild DL, Bonin M, Reuther J, Wohlleben W, Smith MC, Burroughs NJ, Martín JF, Hodgson DA, Takano E, Breitling R, Ellingsen TE, Wellington EM (2010) The dynamic architecture of the metabolic switch in Streptomyces coelicolor. BMC Genomics 11:10–19

    Article  PubMed  Google Scholar 

  • Ryding NJ, Anderson TB, Champness WC (2002) Regulation of the Streptomyces coelicolor calcium-dependent antibiotic by absA, encoding a cluster-linked two-component system. J Bacteriol 184:794–805

    Article  PubMed  CAS  Google Scholar 

  • Sambrook JFE, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor, New York

    Google Scholar 

  • Santos-Beneit F, Rodríguez-García A, Franco-Domínguez E, Martín JF (2008) Phosphate-dependent regulation of the low- and high-affinity transport systems in the model actinomycete Streptomyces coelicolor. Microbiology 154:2356–2370

    Article  PubMed  CAS  Google Scholar 

  • Santos-Beneit F, Rodríguez-García A, Sola-Landa A, Martín JF (2009) Cross-talk between two global regulators in Streptomyces: PhoP and AfsR interact in the control of afsS, pstS and phoRP transcription. Mol Microbiol 72:53–68

    Article  PubMed  CAS  Google Scholar 

  • Santos-Beneit F, Barriuso-Iglesias M, Fernández-Martínez LT, Martínez-Castro M, Sola-Landa A, Rodríguez-García A, Martín JF (2011) The RNA polymerase omega factor RpoZ is regulated by PhoP and has an important role in antibiotic biosynthesis and morphological differentiation in Streptomyces coelicolor. Appl Environ Microbial 77:7586–7594

    Article  CAS  Google Scholar 

  • Sharma CM, Hoffmann S, Darfeuille F, Reignier J, Findeiss S, Sittka A, Chabas S, Reiche K, Hackermüller J, Reinhardt R, Stadler PF, Vogel J (2010) The primary transcriptome of the major human pathogen Helicobacter pylori. Nature 464:250–255

    Article  PubMed  CAS  Google Scholar 

  • Sheeler NL, MacMillan SV, Nodwell JR (2005) Biochemical activities of the absA two-component system of Streptomyces coelicolor. J Bacteriol 187:687–696

    Article  PubMed  CAS  Google Scholar 

  • Sola-Landa A, Rodríguez-García A, Franco-Domínguez E, Martín JF (2005) Binding of PhoP to promoters of Pi-regulated genes in Streptomyces coelicolor: identification of PHO boxes. Mol Microbiol 56:1373–1385

    Article  PubMed  CAS  Google Scholar 

  • Strohl WR (1992) Compilation and analysis of DNA sequences associated with apparent streptomycete promoters. Nucleic Acids Res 20:961–974

    Article  PubMed  CAS  Google Scholar 

  • Thomas L, Hodgson DA, Wentzel A, Nieselt K, Ellingsen TE, Moore J, Morrissey ER, Legaie R, The STREAM Consortium, Wohlleben W, Rodríguez-García A, Martín JF, Burroughs NJ, Wellington EM, Smith MC (2012) Metabolic switches and adaptations deduced from the proteomes of Streptomyces coelicolor wild type and phoP mutant grown in batch culture. Mol Cell Proteomics 11:M111.013797

  • Wayne KJ, Sham LT, Tsui HCT, Gutu AD, Barendt SM, Keen SK, Winkler ME (2010) Localization and cellular amounts of the WalRKJ (VicRKX) two-component regulatory system proteins in serotype 2 Streptococcus pneumoniae. J Bacteriol 192:4388–4394

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This article was supported by grant BIO2010-16094 of the CICYT (Ministry of Science and Innovation, Spain).

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Authors and Affiliations

  1. Instituto de Biotecnología de León, INBIOTEC, Parque Científico de León, Av. Real, 1, 24006, León, Spain

    Fernando Santos-Beneit, Antonio Rodríguez-García & Juan F. Martín

  2. Área de Microbiología, Fac. CC. Biológicas y Ambientales, Universidad de León, Campus de Vegazana, s/n, 24071, León, Spain

    Antonio Rodríguez-García & Juan F. Martín

Authors
  1. Fernando Santos-Beneit
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  2. Antonio Rodríguez-García
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  3. Juan F. Martín
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Correspondence to Juan F. Martín.

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Communicated by G. Klug.

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Santos-Beneit, F., Rodríguez-García, A. & Martín, J.F. Identification of different promoters in the absA1–absA2 two-component system, a negative regulator of antibiotic production in Streptomyces coelicolor . Mol Genet Genomics 288, 39–48 (2013). https://doi.org/10.1007/s00438-012-0728-2

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  • DOI: https://doi.org/10.1007/s00438-012-0728-2

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