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Tandem promoters, tsrp1 and tsrp2, direct transcription of the thiostrepton resistance gene (tsr) of Streptomyces azureus: Transcriptional initiation from tsrp2 occurs after deletion of the — 35 region

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Summary

Nuclease S1 protection experiments indicated that the thiostrepton resistance gene (tsr) of Streptomyces azureus is transcribed from tandem promoters, tsrp1 and tsrp2, that initiate transcription 45 and 173 nucleotides, respectively, upstream of the presumptive translational start codon. The −10 regions of both promoters show similarity to the consensus sequence for the major class of prokaryotic promoters, but the −35 regions do not, although they show some similarity to each other. Replacement of sequences upstream of position −22 relative to the tsrp2 start site with two different DNA segments affected the levels of the tsrp2 transcript but did not alter the tsrp2 initiation site. In vitro transcription assays using RNA polymerase from Streptomyces coelicolor A3(2) also confirmed the location of tsrp2 and identified additional start sites near tsrp2 that were barely detectable with in vivo synthesised RNA. Transcripts corresponding to initiation in vitro at tsrp1 could not be detected, suggesting that additional factors are required for utilisation of this promoter.

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References

  • Baum EZ, Buttner MJ, Lin L-S, Rothstein DM (1989) The P1 promoters of Micromonospora echinospora do not require extensive upstream DNA sequences. J Bacteriol 171:6503–6510

    Google Scholar 

  • Baylis HA, Bibb MJ (1988) Transcriptional analysis of the 16S rRNA gene of the rrnD gene set of Streptomyces coelicolor A3(2). Mol Microbiol 2:569–579

    Google Scholar 

  • Berdy J (1980) Recent advances in and prospects of antibiotic research. Process Biochem Oct/Nov: 28–35

  • Bibb MJ, Janssen GR (1987) Unusual features of transcription and translation of antibiotic resistance genes in antibiotic-producing Streptomyces. In: Alacevic M, Hranueli D, Toman Z (eds) Genetics of Industrial Microorganisms, Part B. Pliva, Zagreb, pp 309–318

    Google Scholar 

  • Bibb MJ, Bibb MJ, Ward JM, Cohen SN (1985) Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces. Mol Gen Genet 199:26–36

    Google Scholar 

  • Bibb MJ, Janssen GR, Ward JM (1986) Cloning and analysis of the promoter region of the erythromycin resistance gene (ermE) of Streptomyces erythraeus. Gene 41:E357-E368

    Google Scholar 

  • Buck M (1986) Deletion analysis of the Klebsiella pneumoniae nitrogenase promoter: Importance of spacing between conserved sequences around positions −12 and −24 for activation by the nifA and ntrC (glnA) products. J Bacteriol 166:545–551

    Google Scholar 

  • Buttner MJ (1989) RNA polymerase heterogeneity in Streptomyces. Mol Microbiol 3:1653–1659

    Google Scholar 

  • Buttner MJ, Brown NL (1985) RNA polymerase-DNA interactions in Streptomyces. In vitro studies of a S. lividans plasmid promoter with S. coelicolor RNA polymerase. J Mol Biol 185:177–188

    Google Scholar 

  • Buttner MJ, Fearnley IM, Bibb MJ (1987) The agarase gene (dagA) of Streptomyces coelicolor A3(2): nucleotide sequence and transcriptional analysis. Mol Gen Genet 209:101–109

    Google Scholar 

  • Buttner MJ, Smith AM, Bibb MJ (1988) At least three different RNA polymerase holoenzymes direct transcription of the agarase gene (dagA) of Streptomyces coelicolor A3(2). Cell 52:599–607

    Google Scholar 

  • Chater KF (1984) Morphological and physiological differentiation in Streptomyces. In: Losick R, Shapiro L (eds) Microbial Development. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 89–115

    Google Scholar 

  • Chater KF, Bruton CJ, Plaskitt KA, Buttner MJ, Mendez C, Helmann JD (1989) The developmental fate of Streptomyces coelicolor hyphae depends crucially on a gene product homologous with the motility σ factor of Bacillus subtilis. Cell 59:133–143

    Google Scholar 

  • Distler J, Ebert A, Mansouri K, Pissowotzki K, Stockmann M, Piepersberg W (1987) Gene cluster for streptomycin biosynthesis in Streptomyces griseus: nucleotide sequence of three genes and analysis of transcriptional activity. Nucleic Acids Res 15:8041–8056

    Google Scholar 

  • Elliott T, Geiduschek EP (1984) Defining a bacteriophage T4 late promoter: absence of a “ −35 region”. Cell 36:211–219

    Google Scholar 

  • Favaloro J, Triesman R, Kamen R (1980) Transcription maps of polyoma virus-specific RNA: Analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol 65:718–749

    Google Scholar 

  • Floss H (1988) Biosynthesis of the antibiotics nosiheptide and thiostrepton. In: Okami Y, Beppu T, Ogawara H (eds) Biology of Actinomycetes '88. Japan Scientific Press, Tokyo, Japan, pp 401–405

    Google Scholar 

  • Fornwald JA, Schmidt FJ, Adams CW, Rosenberg M, Brawner ME (1987) Two promoters, one inducible and one constitutive, control transcription of the Streptomyces lividans galactose operon. Proc Natl Acad Sci USA 84:2130–2134

    Google Scholar 

  • Hawley DK, McClure WR (1983) Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255

    Google Scholar 

  • Hentschel C, Irminger J-C, Bucher P, Birnstiel ML (1980) Sea urchin histone mRNA termini are located in gene regions downstream from putative regulatory sequences. Nature 285:147–151

    Google Scholar 

  • Hopwood DA, Kieser T, Wright HM, Bibb MJ (1983) Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol 129:2257–2269

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces: A laboratory manual. John Innes Foundation, Norwich, UK

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Janssen GR, Malpartida F, Smith CP (1986a) Regulation of gene expression in antibiotic-producing Streptomyces. In: Booth IR, Higgins CF (eds) Regulation of gene expression — 25 years on, 39th Symposium of the Society for General Microbiology. Cambridge University Press, Cambridge, pp 251–276

    Google Scholar 

  • Hopwood DA, Malpartida F, Chater KF (1986b) Gene cloning to analyse the organisation and expression of antibiotic biosynthesis genes in Streptomyces. In: Kleinkauf H, Dohren HV, Dornauer H, Nesemann G (eds) Regulation of secondary metabolite formation, VCH, Weinheim, Federal Republic of Germany, pp 23–33

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Kieser T (1987) Plasmid and phage vectors for gene cloning and analysis in Streptomyces. Methods Enzymol 153:116–166

    Google Scholar 

  • Hoshiko S, Makabe O, Nojiri C, Katsumato K, Satoh E, Nagaoka K (1987) Molecular cloning and characterisation of the Streptomyces hygroscopicus α-amylase gene. J Bacteriol 169:1029–1036

    Google Scholar 

  • Janssen GR, Bibb MJ (1988) Tandem promoters transcribe the thiostrepton resistance gene from Streptomyces azureus and the viomycin resistance gene from Streptomyces vinaceus. In: Okami Y, Beppu T, Ogawara H (eds) Biology of Actinomycetes '88. Japan Scientific Societies Press, Tokyo, Japan pp 374–379

    Google Scholar 

  • Janssen GR, Ward JM, Bibb MJ (1989) Unusual transcriptional and translational features of the aminoglycoside phosphotransferase gene (aph) from Streptomyces fradiae. Genes Dev 3:415–429

    Google Scholar 

  • Kendall KJ, Cohen SN (1988) Complete nucleotide sequence of the Streptomyces lividans plasmid pIJ101 and correlation of the sequence with genetic properties. J Bacteriol 170:4634–4651

    Google Scholar 

  • Keilty S, Rosenberg M (1987) Constitutive function of a positively regulated promoter reveals new sequences essential for activity. J Biol Chem 262:6389–6395

    Google Scholar 

  • Kieser T, Hopwood DA, Wright HM, Thompson CJ (1982) pIJ101, a multi-copy broad host range Streptomyces plasmid: Functional analysis and development of DNA cloning vectors. Mol Gen Genet 185:223–238

    Google Scholar 

  • Lamond AI, Travers AA (1983) Requirement for an upstream element for optimal transcription of a bacterial tRNA gene. Nature 305:248–250

    Google Scholar 

  • Lomovskaya ND, Mkrtumian NM, Danilenko VN (1972) Characterisation of temperate phage ϕC31 isolated from Streptomyces coelicolor A3(2). J Virol 9:258–262

    Google Scholar 

  • Lund PA, Brown NL (1989) Regulation of transcription in Escherichia coli from the mer and merR promoter in the transposon Tn501. J Mol Biol 205:343–353

    Google Scholar 

  • Martin JF, Demain AL (1980) Control of antibiotic synthesis. Microbiol Rev 44:230–252

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base specific chemical cleavages. Methods Enzymol 65:449–560

    PubMed  Google Scholar 

  • McClure WR (1985) Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem 54:171–204

    Google Scholar 

  • Ponnambalam S, Chan B, Busby S (1988) Functional analysis of different sequence elements in the Escherichia coli galactose operon P2 promoter. Mol Microbiol 2:165–172

    Google Scholar 

  • Raibaud O, Schwartz M (1984) Positive control of transcription initiation in bacteria. Annu Rev Genet 18:187–206

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Seno E, Baltz RH (1989) Structural organisation and regulation of antibiotic biosynthesis and resistance genes in actinomycetes. In: Shapiro S (ed) Regulation of secondary metabolism in actinomycetes. CRC Press, Inc., Boca Raton, Florida, USA, pp 1–48

    Google Scholar 

  • Smith CP, Chater KF (1988) Structure and regulation of controlling sequences for the Streptomyces coelicolor glycerol operon. J Mol Biol 204:569–580

    Google Scholar 

  • Sutcliffe JG (1979) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90

    Google Scholar 

  • Tanaka K, Shiina T, Takahashi H (1988) Multiple principal σ factor homologs in eubacteria: identification of the rpoD box. Science 242:1040–1042

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1980) DNA cloning in Streptomyces: resistance genes from antibiotic-producing species. Nature 286:525–527

    Google Scholar 

  • Thompson CJ, Kieser T, Ward JM, Hopwood DA (1982a) Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene 20:51–62

    Article  CAS  PubMed  Google Scholar 

  • Thompson CJ, Skinner RH, Thompson J, Ward JM, Hopwood DA, Cundliffe E (1982b) Biochemical characterization of resistance determinants cloned from antibiotic-producing streptomycetes. J Bacteriol 151:678–685

    Google Scholar 

  • Vögtli M, Hütter R (1987) Characterization of the hydroxystreptomycin phosphotransferase gene (sph) of Streptomyces glaucescens: nucleotide sequence and promoter analysis. Mol Gen Genet 208:195–203

    Google Scholar 

  • Watson JD, Hopkins NH, Roberts JW, Steitz JA, Weiner AM (1987) In: Molecular biology of the gene, 4th edition, vol 1. The Bejamin Cummings Publishing Company, Menlo Park, California, USA, pp 370

    Google Scholar 

  • Westpheling J, Ranes M, Losick R (1985) RNA polymerase heterogeneity in Streptomyces coelicolor. Nature 313:22–27

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene 33:103–119

    Article  CAS  PubMed  Google Scholar 

  • Yu X-M, Reznikoff WS (1986) Deletion analysis of RNA polymerase interaction sites in the Escherichia coli lactose operon regulatory region. J Mol Biol 188:545–553

    Google Scholar 

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  1. Gary R. Janssen

    Present address: Department of Biology, University of Indiana, 47401, Bloomington, Indiana, USA

Authors and Affiliations

  1. John Innes Centre for Plant Science Research, John Innes Institute, NR4 7UH, Colney Lane, Norwich, UK

    Gary R. Janssen & Mervyn J. Bibb

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  1. Gary R. Janssen
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  2. Mervyn J. Bibb
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Communicated H. Hennecke

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Janssen, G.R., Bibb, M.J. Tandem promoters, tsrp1 and tsrp2, direct transcription of the thiostrepton resistance gene (tsr) of Streptomyces azureus: Transcriptional initiation from tsrp2 occurs after deletion of the — 35 region. Mol Gen Genet 221, 339–346 (1990). https://doi.org/10.1007/BF00259397

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  • DOI: https://doi.org/10.1007/BF00259397

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