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Cloning, nucleotide sequence, and transcriptional analysis of the nusG gene of Streptomyces coelicolor A3(2), which encodes a putative transcriptional antiterminator

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Abstract

A 3 kb genomic fragment containing the nusG gene of Streptomyces coelicolor A3(2) was identified, cloned and sequenced. Sequence analysis revealed 3 complete and 2 truncated open reading frames (ORFs): truncated ORF (similar to a Bacillus gene encoding a thermostable aspartate aminotransferase)-secE (94 amino acids; 79.0% similarity to Escherichia coli SecE)-nusG (300 amino acids; 73.3% similarity to E. coli NusG)-rplK (144 amino acids; 88.5% similarity to E. coli ribosomal subunit L11)-truncated rplA (similar to E. coli ribosomal subunit L1). The gene organization secE-nusG-rplKA exactly matches that in E. coli. Transcriptional analyses by the primer extension method revealed one transcriptional start site each for secE and nusG, and two sites for rplK. The presence of promoters was also confirmed with the aid of a promoter-probe vector.

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References

  • Arndt E, Weigel C (1990) Nucleotide sequence of the genes encoding the L11, L1, L10 and L12 equivalent ribosomal proteins from the archaebacterium Halobacterium marismortui. Nucleic Acid Res 18:1285

    Article  CAS  Google Scholar 

  • Chater KF, Hopwood DA (1984) Streptomyces genetics. In: The biology of the actinomycetes. Academic Press, Inc., London, pp 229–286

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  CAS  Google Scholar 

  • Downing WL, Sullivan SL, Gottesman ME, Dennis PP (1990) Sequence and transcription pattern of the essential Escherichia coli secE-nusG operon. J Bacteriol 172:1621–1627

    Article  CAS  Google Scholar 

  • Greenblatt J, Nodwell JR, Mason SW (1993) Transcriptional antitermination. Nature 364:401–406

    Article  CAS  Google Scholar 

  • Heinrich T, Schroder W, Erdmann VA, Hartmann RK (1992) Identification of the gene encoding transcription factor NusG of Thermus thermophilus. J Bacteriol 174:7859–7863

    Article  CAS  Google Scholar 

  • Hillemann D, Pühler A, Wohlleben W (1991) Gene disruption and gene replacement in Streptomyces via single stranded DNA transformation of integration vectors. Nucleic Acids Res 19:727–731

    Article  CAS  Google Scholar 

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

    Google Scholar 

  • Innis MA, Gelfand DH, Sninsky JJ, White JT (1990) PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp 54–59

    Google Scholar 

  • Jeong SM, Yoshikawa H, Takahashi H (1993) Isolation and characterization of the secE homologue gene of Bacillus subtilis. Mol Microbiol 10:133–142

    Article  CAS  Google Scholar 

  • Malpartida F, Hallam SE, Kieser HM, Motamedi H, Hutchinson CR, Butler MJ, Sugden DA, Warren M, McKillop C, Bailey CR, Humphreys GO, Hopwood DA (1987) Homology between Streptomyces genes coding for synthesis of different polyketides used to clone antibiotic synthesis genes. Nature (London) 325:818–821

    Article  CAS  Google Scholar 

  • McConlogue L, Brow MAD, Innis MA (1988) Structure-independent DNA amplification by PCR using 7-deaza-2′-deoxyguanosine. Nucleic Acids Res 16:9869

    Article  CAS  Google Scholar 

  • McKnight SL, Kingsbury R (1982) Transcriptional control signals of eukaryotic protein-encoding gene. Science 217:316–324

    Article  CAS  Google Scholar 

  • Miyake K, Onaka H, Horinouchi S, Beppu T (1994) Organization and nucleotide sequence of the secE-nusG region of Streptomyces griseus. Biochim Biophys Acta 1217:97–100

    Article  CAS  Google Scholar 

  • Muth G, Nußbaumer B, Wohlleben W, Pühler A (1989) A vector system with temperature-sensitive replication for gene disruption and mutational cloning in Streptomyces. Mol Gen Genet 219:341–348

    Article  CAS  Google Scholar 

  • Okamoto S, Nihira T, Kataoka H, Suzuki A, Yamada Y (1992) Purification and molecular cloning of a butyrolactone autoregulator receptor from Streptomyces virginiae. J Biol Chem 267:1093–1098

    CAS  PubMed  Google Scholar 

  • Post LE, Strycharz GD, Nomura M, Lewis H, Dennis PP (1979) Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit β in Escherichia coli. Proc Natl Acad Sci USA 76:1697–1701

    Article  CAS  Google Scholar 

  • Puttikhunt C, Okamoto S, Nakamura T, Nihira T, Yamada Y (1993) Distribution in the genus Streptomyces of a homolog to nusG, a gene encoding a transcriptional antiterminator. FEMS Microbiol Lett 110:243–248

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, A laboratory manual, 2nd edition, Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Schatz PJ, Bieker KL, Ottemann KM, Silhavy TJ, Beckwith J (1991) One of three transmembrane stretches is sufficient for the functioning of the SecE protein, a membrane component of the E. coli secretion machinery. EMBO J 10:1749–1757

    Article  CAS  Google Scholar 

  • Schatz PJ, Riggs PD, Jacq A, Fath MJ, Beckwith J (1989) The secE gene encodes an integral membrane protein required for protein export in E. coli. Genes Dev 3:1035–1044

    Article  CAS  Google Scholar 

  • Shimmin LC, Dennis PP (1989) Characterization of the L11, L1, L10 and L12 equivalent ribosomal protein gene cluster of the halophilic archaebacterium Halobacterium cutirubrum. EMBO J 8:1225–1235

    Article  CAS  Google Scholar 

  • Sor F, Nomura M (1987) Cloning and DNA sequence determination of the L11 ribosomal protein operon of Serratia marcescens and Proteus vulgaris: Translational feedback regulation of the Escherichia coli L11 operon by heterologous L1 proteins. Mol Gen Genet 210:52–59

    Article  CAS  Google Scholar 

  • Sung MH, Tanizawa K, Tanaka H, Kuramitsu S, Kagamiyama H, Hirotsu K, Okamoto A, Higuchi T, Soda K (1991) Thermostable aspartate aminotransferase from a thermophilic Bacillus species — Gene cloning, sequence determination and preliminary X-ray characterization. J Biol Chem 266:2567–2572

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33:103–119

    Article  CAS  Google Scholar 

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Communicated by J. F. Lengeler

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Puttikhunt, C., Nihira, T. & Yamada, Y. Cloning, nucleotide sequence, and transcriptional analysis of the nusG gene of Streptomyces coelicolor A3(2), which encodes a putative transcriptional antiterminator. Molec. Gen. Genet. 247, 118–122 (1995). https://doi.org/10.1007/BF00425829

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