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Cloning and DNA sequence analysis of the mercury resistance genes of Streptomyces lividans

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Summary

A broad-spectrum mercury resistance locus (mer) from a spontaneous chloramphenicol-sensitive (Cms), arginine auxotrophic (Arg−) mutant of Streptomyces lividan 1326 was isolated on a 6 kb DNA fragment by shotgun cloning into the mercury-sensitive derivative S. lividans TK64 using the vector pIJ702. The mer genes form part of a very large amplifiable DNA sequence present in S. lividans 1326. This element was amplified to about 20 copies per chromosome in the Cms Arg− mutant and was missing from strains like S. lividans TK64, cured for the plasmid SLP3. DNA sequence analysis of a 5 kb region encompassing the whole region required for broad-spectrum mercury resistance revealed six open reading frames (ORFs) transcribed in opposite directions from a common intercistronic region. The protein sequences predicted from the two ORFs transcribed in one direction showed a high degree of similarity to mercuric reductase and organomercurial lyase from other gram-negative and gram-positive sources. Few, if any, similarities were found between the predicted polypeptide sequences of the other four ORFs and other known proteins.

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References

  • Altenbuchner J, Cullum J (1985) Structure of an amplifiable DNA sequence in Streptomyces lividans 66. Mol Gen Genet 201:192–197

    Google Scholar 

  • Babich K, Engle JM, Skinner JS, Laddaga RA (1991) Deletion mutant analysis of the Staphylococcus aureus plasmid pI258 mercury resistance determinant. Canad J Microbiol 37:624–631

    Google Scholar 

  • Bernan V, Filpula D, Herber W, Bibb MJ, Katz E (1985) The nucleotide sequence of the tyrosinase gene from Streptomyces antibioticus and characterisation of the gene product. Gene 37:101–110

    Google Scholar 

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

    Google Scholar 

  • Bibb MJ, Ward JH, Cohen SN (1985) Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces. Mol Gen Genet 199:26–39

    Google Scholar 

  • Bibb MJ (1986) Gene expression in Streptomyces: nucleotide sequences involved in initiation of transcription and translation. In: Szabó G, Biro S, Goodfellow M (eds) Biological, biochemical and biomedical aspects of Actinomycetes, Part A. Akadémiai Kiadó, Budapest

    Google Scholar 

  • Bogdanova ES, Mindlin SZ (1989) Two structural types of mercury reductases and possible ways of their evolution. FEBS Lett 247:333–336

    Google Scholar 

  • Brown NL, Ford SJ, Pridmore RD, Fritzinger DC (1983) Nucleotide sequence of a gene from the Pseudomonas transposon TN501 encoding mercuric reductase. Biochemistry 22:4089–4095

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Eichenseer C, Altenbuchner J (1991) Amplifiable elements of Streptomyces lividans are useful tools to increase gene expression. In: Reuss M, Chmiel H, Gilles E-D, Knackmuss H-J (eds) Biochemical engineering-Stuttgart, Gustav Fischer, Stuttgart

    Google Scholar 

  • Fox BS, Walsh CT (1983) Mercuric reductase: homology to glutathione reductase and lipoamide dehydrogenase. Iodoacetamide alkylation and sequence of the active site peptide. Biochemistry 22:4082–4088

    Google Scholar 

  • Gambill BD, Summers AO (1985) Versatile mercury-resistant cloning and expression vectors. Gene 39:293–297

    Google Scholar 

  • Gribskov M, Devereux J, Burgess RR (1984) The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res 12:539–549

    Google Scholar 

  • Griffin HG, Foster TJ, Silver S, Misra TK (1987) Cloning and DNA sequence of the mercuric and organomercurial resistance determinants of plasmid pDU1358. Proc Natl Acad Sci USA 84:3112–3116

    Google Scholar 

  • Henderson G, Krygsman P, Jun Liu CI, Davey CC, Malek LT (1987) Characterization and structure of genes for proteases A and B from Streptomyces griseus. J Bacteriol 169:3778–3784

    Google Scholar 

  • Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

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

    Google Scholar 

  • Hopwood DA, Lydiate DJ, Malpartida F, Wright HM (1984) Conjugative sex plasmids in Streptomyces. In: Helinski D, Cohen S, Clewell D (eds) Plasmids in bacteria. Plenum Press, New York, pp 615–634

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater RF, 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 

  • Hussain M, Pastor FIJ, Lampen JO (1987) Cloning and sequencing of the blaZ gene encoding β-lactamase III, a lipoprotein of Bacillus cereus 569/H. J Bacteriol 169:579–586

    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 

  • Kieser T (1984) Factors affecting the isolation of ccc-DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36

    Google Scholar 

  • Kusano T, Jl G, Inoue C, Silver S (1990) Constitutive synthesis of a transport function encoded by the Thiobacillus ferrooxidans merC gene cloned in Escherichia coli. J Bacteriol 172:2688–2692

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. Nucleic Acids Res 157:105–132

    Google Scholar 

  • Laddaga RA, Chu L, Misra TK, Silver S (1987) Nucleotide sequence and expression of the mercurial resistance operon from Staphylococcus aureus plasmid p1258. Proc Natl Acad Sci USA 84:5106–5110

    Google Scholar 

  • Marsh LJ, Erfle M, Wykes EJ (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32:481–485

    Google Scholar 

  • McKenney R, Shimatake H, Court D, Schmeissner U, Braty C, Rosenberg M (1981) Analysis of nucleic acids by enzymatic methods. In: Chirikyian JG, Papas TS (eds), Gene amplification and analysis, Vol. 2, Elsevier North-Holland, Amsterdam, pp 383–415

    Google Scholar 

  • Misra TK (1992) Bacterial resistances to inorganic mercury salts and organomercurials. Plasmid 27:4–16

    Google Scholar 

  • Misra TK, Brown N, Haberstroh L, Schmidt A, Godette D, Silver S (1985) Mercuric reductase structural genes from plasmid R100 and transposon Tn501: functional domains of the enzyme. Gene 34:253–262

    Google Scholar 

  • Nakahara H, Silver S, Miki T, Rownd RH (1979) Hypersensitivity to Hg2+ and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of plasmid NR1. J Bacteriol 140:161–166

    Google Scholar 

  • Nakahara H, Schottel JL, Yamada T, Miyakawa Y, Asakawa M, Harville J, Silver S (1985) Mercuric reductase enzymes from Streptomyces species and Group B Streptococcus. J Gen Microbiol 131:1053–1059

    Google Scholar 

  • O'Halloran T, Frantz B, Shin MK, Ralston DM, Wright JC (1989) The merR metal receptor mediates positive activation in a topologically novel transcription complex. Cell 56:119–129

    Google Scholar 

  • Polsinelli M, Beretta M (1966) Genetic recombination in crosses between Streptomyces aureofaciens and Streptomyces rimosus. J Bacteriol 91:63–68

    Google Scholar 

  • Robbins PW, Trimble RB, Wirth DF, Hering F, Maley GF, Maley RDas, Gibson BW, Royal N, Bieman K (1984) Primary structure of the Streptomyces enzyme endo-β-N-acetylglucosaminidase H. J Biol Chem 259:7577–7583

    Google Scholar 

  • Rosenstein R, Peschel A, Wieland B, Götz F (1992) Expression and regulation of the antimony, arsenite and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267. J Bacteriol 174:3676–3683

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • San Francisco MDJ, Hope CL, Owolabi JB, Tisa LS, Rosen BP (1990) Identification of the metalloregulatory element of the plasmid-encoded arsenical resistance operon. Nucleic Acids Res 18:619–624

    Google Scholar 

  • Silver S, Misra TK, Laddaga A (1989) DNA sequence analysis of bacterial toxic heavy metal resistances. Biol Trace Element Res 21:145–163

    Google Scholar 

  • Strohl WR (1922) Compilation and analysis of DNA sequences associated with streptomycete promoters. Nucleic Acids Res 20:961–974

    Google Scholar 

  • Tautz D, Renz M (1983) An optimized freeze-squeeze methode for the recovery of DNA fragments from agarose gels. Anal Biochem 132:14–19

    Google Scholar 

  • Tohyama H, Okami Y, Umezawa H. (1987) Nucleotide sequence of the streptomycinphosphotransferase and amidonotransferase genes from Streptomyces griseus. Nucl Acids Res 15:1819–1833

    Google Scholar 

  • van Heijne G (1989) The structure of signal peptides from bacterial lipoproteins. Protein Eng 2:531–534

    Google Scholar 

  • Wang Y, Moore M, Levinson HS, Silver S, Walsh C, Mahler I (1989) Nucleotide sequence of a chromosomal mercury resistance determinant from a Bacillus sp. with broad-spectrum mercury resistance. J Bacteriol 171:83–92

    Google Scholar 

  • Walsh CT, Distefano MD, Moore MJ, Shewchuk LM, Verdine GL (1988) Molecular basis of bacterial resistance to organomercurial and inorganic mercury salts. FEBS 2:124–130

    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

    Google Scholar 

  • Zagursky R, Baumeister K, Lomax N, Berman M (1986) Dideoxy DNA sequencing directly from plasmid DNA using reverse transcriptase. Gene Anal Tech 2:89–95

    Google Scholar 

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Authors and Affiliations

  1. Institut für Genetik und Mikrobiologie, Universität München, Maria Ward Strasse IA, W-8000, München 19, Germany

    Reinhard Sedlmeier

  2. Institut für industrielle Genetik, Universität Stuttgart, Azenbergstrasse 18, W-7000, Stuttgart 1, Germany

    Josef Altenbuchner

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  1. Reinhard Sedlmeier
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  2. Josef Altenbuchner
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Communicated by J. Lengeler

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Sedlmeier, R., Altenbuchner, J. Cloning and DNA sequence analysis of the mercury resistance genes of Streptomyces lividans . Molec. Gen. Genet. 236, 76–85 (1992). https://doi.org/10.1007/BF00279645

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  • DOI: https://doi.org/10.1007/BF00279645

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