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The agarase gene (dagA) of Streptomyces coelicolor A3(2): nucleotide sequence and transcriptional analysis

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Summary

The DNA sequence of a 1.77 kb region of the Streptomyces coelicolor chromosome containing the coding and regulatory regions of the extracellular agarase (dagA) gene was determined. The sequence predicts a primary translation product of 309 amino acids and Mr 35132. Comparison of the N-terminal sequence determined for the mature extracellular protein with that of the primary translation product deduced from the DNA sequence predicts the presence of a 30 amino acid signal peptide. Analysis of the transcription of the dagA gene using high resolution S1 mapping, in vitro transcription, dinucleotide-primed in vitro transcription and in vivo promoter probing identified four promoters, initiating transcription approximately 32, 77, 125 and 220 nucleotides upstream of the coding sequence.

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References

  • Abrahmsén L, Moks T, Nilsson B, Hellman U, Uhlén M (1985) Analysis of signals for secretion in the staphylococcal protein A gene. EMBO J 4:3901–3906

    Google Scholar 

  • Adhya S, Garges S (1982) How cyclic AMP and its receptor protein act in Escherichia coli. Cell 29:287–289

    Google Scholar 

  • Bankier AT, Barrell BG (1983) Shotgun DNA sequencing. In: Flavell RA (ed) Techniques in the life sciences. Elsevier Press, Ireland, pp 1–34

    Google Scholar 

  • Benigni R, Antonov RP, Carere A (1975) Estimate of the genome size by renaturation studies in Streptomyces. Appl Microbiol 30:324–326

    Google Scholar 

  • Bibb MJ, Cohen SN (1982) Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet 187:265–277

    Google Scholar 

  • Bibb MJ, Janssen GR (1987) Unusual features of transcription and translation of antibiotic resistance genes in two antibioticproducing Streptomyces species. Proceedings of the 5th International Conference of the Genetics of Industrial Micro-organisms. Split, Yugoslavia (in press)

  • Bibb MJ, Freeman RF, Hopwood DA (1977) Physical and genetical characterisation of a second sex factor, SCP2, for Streptomyces coelicolor. Mol Gen Genet 154:155–166

    Google Scholar 

  • Bibb MJ, Findlay PR, Johnson MW (1984) The relationship between base composition and codon usage in bacterial genes and its use in the simple and reliable identification of protein coding sequences. Gene 30:157–166

    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 

  • Bibb MJ, Jones GH, Joseph R, Buttner MJ, Ward JM (1987) The agarase gene (dagA) of Streptomyces coelicolor A3(2): affinity purification and characterisation of the cloned gene product. J Gen Microbiol, in press

  • Botsford JL (1981) Cyclic nucleotides in procaryotes. Microbiol Rev 45:620–642

    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 

  • Chang S (1987) Engineering for protein secretion in Gram-positive bacteria. Methods Enzymol (in press)

  • Chatterjee S, Vining LC (1982) Catabolite repression in Streptomyces venezuelae. Induction of β-galactosidase, chloramphenicol production and intracellular cyclic adenosine 3′–5′-monophosphate concentrations. Can J Microbiol 28:311–317

    Google Scholar 

  • Cortes J, Liras P, Castro JM, Martín JF (1986) Glucose regulation of cephamycin biosynthesis in Streptomyces lactamdurans is exerted on the formation of α-aminoadipyl-cysteinyl-valine and deacetoxycephalosporin C synthase. J Gen Microbiol 132:1805–1814

    Google Scholar 

  • de Crombrugghe B, Busby S, Buc H (1984) Cyclic AMP, the cyclic AMP receptor protein, and their dual control of the galactose operon. Science 224:831–838

    Google Scholar 

  • Deininger PL (1983) Random subcloning of sonicated DNA: application to shotgun DNA sequence analysis. Anal Biochem 129:216–223

    Google Scholar 

  • Doolittle RF (1981) Similar amino acid sequences: chance or common ancestry? Science 214:149–159

    Google Scholar 

  • Downey KM, Jurmark BS, So AG (1971) Determination of nucleotide sequence at promoter regions by the use of dinucleotides. Biochemistry 10:4970–4975

    Google Scholar 

  • Enquist LW, Bradley SG (1971) Characterisation of deoxyribonucleic acid from Streptomyces venezuelae spores. Dev Ind Microbiol 12:225–236

    Google Scholar 

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

    Google Scholar 

  • Hodgson DA (1982) Glucose repression of carbon source uptake and metabolism in Streptomyces coelicolor A3(2) and its perturbation in mutants resistant to 2-deoxyglucose. J Gen Microbiol 128:2417–2430

    Google Scholar 

  • Hodgson DA, Chater KF (1981) A chromosomal locus controlling extracellular agarase production by Streptomyces coelicolor A3(2) and its inactivation by chromosomal integration of plasmid SCP1. J Gen Microbiol 124:339–348

    Google Scholar 

  • Hoffman DJ, Niyogi SK (1973) RNA initiation with dinucleoside monophosphates during transcription of bacteriophage T4 DNA with RNA polymerase of Escherichia coli. Proc Natl Acad Sci USA 70:574–578

    Google Scholar 

  • Hopwood DA (1967) Genetic analysis and genome structure in Streptomyces coelicolor. Bacteriol Rev 31:373–403

    Google Scholar 

  • Hopwood DA, Chater KF, Dowding JF, Vivian A (1973) Advances in Streptomyces coelicolor genetics. Bacteriol Rev 37:371–405

    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

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Janssen GR, Malpartida F, Smith CP (1986) Regulation of gene expression in autibiotic-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

    Google Scholar 

  • Katz E, Thompson CJ, Hopwood DA (1983) Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol 129:2703–2714

    Google Scholar 

  • Kendall K, Cullum J (1984) Cloning and expression of an extracellular-agarase gene from Streptomyces coelicolor A3(2) in Streptomyces lividans 66. Gene 29:315–321

    Google Scholar 

  • Krakow JS, Rhodes G, Jovin TM (1976) In: Losick R, Chamberlin M (eds) RNA polymerase. Cold Spring Harbor Laboratory Press, New York, pp 127–158

    Google Scholar 

  • Martín JF, Demain AL (1980) Control of antibiotic biosynthesis. Microbiol Rev 44:230–251

    Google Scholar 

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

    Google Scholar 

  • Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either strand of double-digest restriction fragments. Gene 19:269–276

    Google Scholar 

  • Minkley EG, Pribnow D (1973) Transcription of the early region of bacteriophage T7: selective initiation with dinucleotides. J Mol Biol 77:255–277

    Google Scholar 

  • Moran CP Jr, Lang N, Banner CDB, Haldenwang WG, Losick R (1981) Promoter for a developmentally regulated gene in Bacillus subtilis. Cell 25:783–791

    Google Scholar 

  • Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligonucleotide-directed mutagenesis. Gene 26:101–106

    Google Scholar 

  • Perlman D, Halvorson HO (1983) A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol 167:391–409

    Google Scholar 

  • Pugsley AP, Schwartz M (1985) Export and secretion of proteins by bacteria. FEMS Microbiol Rev 32:3–38

    Google Scholar 

  • Sanger F, Coulson AR (1978) The use of thin polyacrylamide gels for DNA sequencing. FEBS Lett 186:107–110

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5468

    Google Scholar 

  • Sanger F, Coulson AR, Barrell BG, Smith AJH, Roe BA (1980) Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′-terminal sequence of E. coli 16S ribosomal RNA: complimentarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Staden R (1982) Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res 10:4731–4751

    Google Scholar 

  • Staden R (1984a) A computer program to enter DNA gel reading data into a computer. Nucleic Acids Res 12:499–503

    Google Scholar 

  • Staden R (1984b) Computer methods to locate signals in nucleic acid sequences. Nucleic Acids Res 12:505–515

    Google Scholar 

  • Staden R (1984c) Graphic methods to determine the function of nucleic acid sequences. Nucleic Acids Res 12:521–538

    Google Scholar 

  • Stanier RY (1942) Agar decomposing strains of the Actinomyces coelicolor species group. J Bacteriol 44:555–570

    Google Scholar 

  • Surowitz KG, Pfister RM (1985) Variations in levels of cAMP, DNA and RNA in Streptomyces alboniger under conditions of aerial mycelium formation and repression. FEMS Microbiol Lett 26:1–4

    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 

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

    Google Scholar 

  • von Heijne G (1984) How signal sequences maintain cleavage specificity. J Mol Biol 173:243–251

    Google Scholar 

  • Ward JM, Janssen GR, Kieser T, Bibb MJ, Buttner MJ, Bibb MJ (1986) Construction and characterisation of a series of multi-copy promoter-probe plasmid vectors for Streptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicator. Mol Gen Genet 203:468–478

    Google Scholar 

  • Watson MEE (1984) Compilation of published signal sequences. Nucleic Acids Res 12:5145–5164

    Google Scholar 

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

    Google Scholar 

  • Wilbur WJ, Lipman DJ (1983) Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci USA 80:726–730

    Google Scholar 

  • Williams ST, Goodfellow M, Alderson G, Wellington EMH, Sneath PHA, Sackin MJ (1983) Numerical classification of Streptomyces and related genera. J Gen Microbiol 129:1743–1813

    Google Scholar 

  • Zalacain M, González A, Guerrero MC, Mattaliano RT, Malpartida F, Jiménez A (1986) Nucleotide sequence of the hygromycin B phosphotransferase gene from Streptomyces hygroscopicus. Nucleic Acids Res 14:1565–1581

    Google Scholar 

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Authors and Affiliations

  1. John Innes Institute, Colney Lane, NR4 7UH, Norwich, UK

    Mark J. Buttner & Mervyn J. Bibb

  2. Medical Research Council Laboratory of Molecular Biology, Medical Research Council Centre, Hills Road, CB2 2QH, Cambridge, UK

    Ian M. Fearnley

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  1. Mark J. Buttner
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  2. Ian M. Fearnley
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  3. Mervyn J. Bibb
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Communicated by W. Goebel

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Buttner, M.J., Fearnley, I.M. & Bibb, M.J. The agarase gene (dagA) of Streptomyces coelicolor A3(2): nucleotide sequence and transcriptional analysis. Mol Gen Genet 209, 101–109 (1987). https://doi.org/10.1007/BF00329843

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  • DOI: https://doi.org/10.1007/BF00329843

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