Skip to main content
SpringerLink
Log in

Genetics of streptomycin production in Streptomyces griseus: molecular structure and putative function of genes strELMB2N

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The nucleotide sequence of a 5.1 kb fragment from the streptomycin biosynthetic gene cluster from Streptomyces griseus revealed the presence of five open reading frames which form part of two convergently oriented transcription units strDEL and strNB2M. The coding capacity for polypeptide products was calculated to be 35.7 kDa (StrE), 32.2 kDa (StrL), 35.6 kDa (StrN), 38.2 kDa (StrB2), and 21.9 kDa (StrM), respectively. Various observations suggested that the gene products StrD (dTDP-glucose synthase), StrE (dTDP-glucose dehydratase), StrM (dTDP-4-keto-6-deoxyglucose 3,5-epimerase), and StrL (dTDP-dihydrostreptose synthase) are involved in biosynthesis of the streptose moiety of streptomycin. StrE and StrL are significantly similar in primary structure to each other and to other oxidoreductases (epimerases) involved in hexose metabolism. Genes for dTDP-glucose synthase and dehydratase occur in other gene clusters for antibiotic production. Therefore, the strD and strE genes could serve as universal probes indicative of the presence of biosynthetic capacity for 6-deoxyhexose moieties. The StrB2 protein showed 69% amino acid identity to the first-step amidinotransferase StrBl. The presence of both strB genes appears to be the result of a gene duplication event. The gene product StrN contains sequence motifs also conserved in the putative catalytic and/or substrate recognition domains of aminoglycoside phosphotransferases and eucaryotic protein kinases. The possible role of a TTA codon, located near the start of the strN reading frame, in regulation of the str cluster is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams CW, Fornwald JA, Schmidt FJ, Rosenberg M, Brawner M (1988) Gene organization and structure of the Streptomyces lividans gal operon. J Bacteriol 170:203–212

    Google Scholar 

  • Ambulos NP, Smith T, Mulbry W, Lovett PS (1990) CUG as a mutant start codon for cat-86 and xylE in Bacillus subtilis. Gene 94:125–128

    Google Scholar 

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

    Google Scholar 

  • Birch A, Häusler A, Vögtli M, Krek W, Hütter R (1989) Extremely large chromosomal deletions are intimately involved in genetic instability and genomic rearrangements in Streptomyces glaucescens. Mol Gen Genet 217:447–458

    Google Scholar 

  • Chater KF (1989) Sporulation in Streptomyces. In: Smith I, Slepecky RA, Setlow P (eds) Regulation of procaryotic development. American Society for Microbiology, Washington, DC, pp 277–299

    Google Scholar 

  • Citron BA, Donelson JE (1984) Sequence of the Saccharomyces GAL region and its transcription in vivo. J Bacteriol 158:269–78

    Google Scholar 

  • Distler J, Klier K, Piendl W, Böck A, Kresze G, Piepersberg W (1985a) Streptomycin biosynthesis in Streptomyces griseus I. Characterization of streptomycin-idiotrophic mutants. FEMS Microbiol Lett 30:145–150

    Google Scholar 

  • Distler J, Mansouri K, Piepersberg W (1985b) Streptomycin biosynthesis in Streptomyces griseus II. Adjacent genomic location of biosynthetic genes and one of two streptomycin resistance genes. FEMS Microbiol Lett 30:151–154

    Google Scholar 

  • Distler J, Braun C, Ebert A, Piepersberg W (1987a) Gene cluster for streptomycin biosynthesis in Streptomyces griseus: Analysis of a central region including the major resistance gene. Mol Gen Genet 208:204–210

    Google Scholar 

  • Distler J, Ebert A, Mansouri K, Pissowotzki K, Stockmann M, Piepersberg W (1987b) 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 

  • Distler J, Mansouri K, Mayer G, Piepersberg W (1990) Regulation of biosynthesis of streptomycin. In: Heslot H, Davies J, Florent J, Bobichon L, Durant G, Penasse L (eds) The proceedings of the sixth international symposium on Genetics of Industrial Microorganisms. Société Francaise de Microbiologie, Strasbourg, pp 379–392

    Google Scholar 

  • Fishbein L, Cavanaugh MA (1965) Detection and paper chromatography of N-substituted hydroxy-, 2-hydroxyethyl-, 2-chloroethyl, and N,N-bis(2-hydroxyethyl)-derivatives. J Chromatogr 20:283–289

    Google Scholar 

  • Gabriel O (1987) Biosynthesis of sugar residues for glycogen, peptidoglycan, lipopolysaccharide, and related systems. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium. Cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 504–511

    Google Scholar 

  • Grisebach H (1978) Biosynthesis of sugar components of antibiotic substances. Adv Carbohydr Chem Biochem 35:81–126

    Google Scholar 

  • Heinzel P, Werbitzky O, Distler J, Piepersberg W (1988), A second streptomycin resistance gene from Streptomyces griseus codes for streptomycin 3″-phosphotransferase. Relationship between antibiotic and protein kinases. Arch Microbiol 150:184–192

    Google Scholar 

  • Hopwood DA, Sherman DH (1990) Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet 24:37–66

    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. The John Innes Foundation, Norwich

    Google Scholar 

  • Hunter IS, Baumberg S (1989) Molecular genetics of antibiotic formation. In: Baumberg S, Hunter IS, Rhodes PM (eds) Microbial products: new approaches. Cambridge University Press, Cambridge, pp 121–162

    Google Scholar 

  • Hutchinson CR, Borell CW, Otten SL, Stutzman-Engwall KJ, Wang Y-G (1989) Drug discovery and development through the genetic engineering of antibiotic-producing microorganisms. J Med Chem 32:929–936

    Google Scholar 

  • Hütter RA, Eckhardt T (1988) Genetic manipulation. In: Goodfellow M, Williams ST, Mordarski M (eds) Actinomycetes in biotechnology. Academic Press, London, pp 89–184

    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 

  • van Knippenberg PH (1990) Aspects of translational initiation in Escherichia coli. In: Hill WE, Dahlberg A, Garrett RA, Moore PB, Schlessinger D, Warner JR (eds) The ribosome — structure, function, and evolution. American Society for Microbiology, Washington, DC, pp 265–274

    Google Scholar 

  • Kumar A, Larsen CE, Preiss J (1986) Biosynthesis of bacterial glycogen. Primary structure of Escherichia coli ADP-glucose: β-1,4-glucan 4-glucosyltransferase as deduced from the nucleotide sequence of the glgA gene. J Biol Chem 261:16256–16259

    Google Scholar 

  • Lee N, Gielow W, Martin R, Hamilton E, Fowler A (1986) The organization of the araBAD operon of Escherichia coli. Gene 47:231–244

    Google Scholar 

  • Lemaire HG, Müller-Hill B (1986) Nucleotide sequence of the galE gene and the galT gene of E. coli. Nucleic Acids Res 14:7705–7711

    Google Scholar 

  • Leskiw BK, Lawlor EJ, Fernandezabalos JM, Chater KF (1991) TTA codons in some genes prevent their expression in a class of developmental, antibiotic-negative, Streptomyces mutants. Proc Natl Acad Sci USA 88:2461–2465

    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 biosynthetic genes. Nature 325:818–821

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Mansouri K, Piepersberg W (1991) Genetics of streptomycin production in Streptomyces griseus: nucleotide sequence of five genes, strFGHIK, including a phosphatase gene. Mol Gen Genet 228:459–469

    Google Scholar 

  • Mansouri K, Pissowotzki K, Distler J, Mayer G, Braun C, Ebert A, Piepersberg W (1989) Genetics of streptomycin production. In: Hershberger CL, Qeener SW, Hegeman G (eds) Genetics and molecular biology of industrial microorganisms. American Society for Microbiology, Washington, DC, pp 61–67

    Google Scholar 

  • Martín JF, Liras P (1989) Organization and expression of genes involved in the biosynthesis of antibiotics and other secondary metabolites. Annu Rev Microbiol 43:173–206

    Google Scholar 

  • Mayer G, Vögtli M, Pissowotzki K, Hütter R, Piepersberg W (1988) Colinearity of streptomycin production genes in two species of Streptomyces. Evidence for occurrence of a second amidinotransferase gene. Mol Genet (Life Sci Adv) 7:83–87

    Google Scholar 

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

    Google Scholar 

  • Ohnuki T, Imanaka T, Aiba S (1985a) Isolation of streptomycin-nonproducing mutants deficient in the biosynthesis of the streptidine moiety or linkage between streptidine 6-phosphate and dihydrostreptose. Antimicrob Agents Chemother 27:367–374

    Google Scholar 

  • Ohnuki T, Imanaka T, Aiba S (1985b) Self-cloning in Streptomyces griseus of an str gene cluster for streptomycin biosynthesis and streptomycin resistance. J Bacteriol 164:85–94

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence analysis. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Piepersberg W, Distler J, Heinzel P, Perez-Gonzalez J-A (1988) Antibiotic resistance by modification: Many resistance genes could be derived from cellular control genes in actinomycetes. — A hypothesis. Actinomycetology 2:83–98

    Google Scholar 

  • Piepersberg W, Heinzel P, Mansouri K, Mönnighoff U, Pissowotzki K (1991) Evolution of antibiotic resistance and production genes in streptomycetes. In: Noack D, Krügel H, Baumberg S (eds) Genetics and product formation in Streptomyces. Plenum Press, London, pp 161–170

    Google Scholar 

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

    Google Scholar 

  • Schürrle K (1991) Enantio- and stereoselektive Darstellungen von cis-1,3-Diamino-1,3-dideoxycyclitolen zu Mutasyntheseexperimenten mit Streptomyces-Mutanten und als Testsubstrate für die Phosphotransferase APH(6) aus S. griseus 40236. Dissertation, Technische Hochschule, Darmstadt, FRG

    Google Scholar 

  • Seno ET, Baltz RH (1989) Structural organization and regulation of antibiotic biosynthesis and resistance genes in actinomycetes. In: Shapiro S (ed) Regulation of secondary metabolism in actinomycetes. CRC Press, London, pp 1–48

    Google Scholar 

  • Thompson CJ, Gray GS (1983) Nucleotide sequence of a streptomycete aminoglycoside phosphotransferase gene and its relationship to phosphotransferases encoded by resistance plasmids. Proc Natl Acad Sci USA 80:5190–5194

    Google Scholar 

  • Tohyama H, Okami Y, Umezawa H (1987) Nucleotide sequence of the streptomycin phosphotransferase and amidinotransferase from Streptomyces griseus. Nucleic Acids Res 15:1819–1834

    Google Scholar 

  • Vara JA, Hutchinson CR (1988) Purification of thymidine-diphospho-D-glucose 4,6-dehydratase from an erythromycin-producing strain of Saccharopolyspora erythrea by high resolution liquid chromatography. J Biol Chem 263:14992–14995

    Google Scholar 

  • Verma N, Reeves P (1989) Identification and sequence of rfbS and rfbE, which determine antigenic specificity of group A and group D salmonellae. J Bacteriol 171:5694–5701

    Google Scholar 

  • Vining LC (1990) Functions of secondary metabolites. Annu Rev Microbiol 44:395–427

    Google Scholar 

  • Vögtli M (1987) Structure and expression of the sph gene of Streptomyces glaucescens and its genomic environment. Dissertation no. 8366, ETH Zürich, Switzerland

    Google Scholar 

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

    Google Scholar 

  • Wahl HP, Grisebach H (1979) Biosynthesis of streptomycin. dTDP-dihydrostreptose synthase from Streptomyces griseus and dTDP-4-keto-L-rhamnose 3,5-epimerase from S. griseus and Escherichia coli Y10. Biochim Biophys Acta 568:243–252

    Google Scholar 

  • Walker JB (1969) Streptomycin biosynthesis: enzymatic synthesis of inosamine and inosadiamine derivatives in the biosynthesis of streptidine. Ann NY Acad Sci 165:646–654

    Google Scholar 

  • Walker JB (1974) Biosynthesis of the monoguanidated inosotol moiety of bluensomycin, a possible evolutionary precursor for streptomycin. J Biol Chem 249:2397–2404

    Google Scholar 

  • Walker JB (1975) Pathways of the guanidinated inositol moieties of streptomycin and bluensomycin. Methods Enzymol 43:429–470

    Google Scholar 

  • Walker JB (1990) Possible evolutionary relationship between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities. J Bacteriol 172:5844–5851

    Google Scholar 

  • Webster TD, Dickson RC (1988) Nucleotide sequence of the galactose gene cluster of Kluyveromyces lactis. Nucleic Acids Res 16:8192

    Google Scholar 

  • Wyk P, Reeves P (1989) Identification and sequence of the gene for abequose synthase, which confers antigenic specificity on group B salmonellae: homology with galactose epimerase. J Bacteriol 171:5687–5693

    Google Scholar 

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

    Google Scholar 

  • Zähner H, Anke H, Anke T (1983) Evolution and secondary pathways. In: Bennett JW, Ciegler A (eds) Secondary metabolism and differentiation in fungi. Marcel Dekker, New York, pp 153–171

    Google Scholar 

Download references

Author information

Author notes

  1. Klaus Pissowotzki

    Present address: , A.F. Pfreudt-Strasse 2, W-6120, Michelstadt, Germany

Authors and Affiliations

  1. Lehrstuhl für Mikrobiologie, Bergische Universität GH, Gauss-Strasse 20, W-5600, Wuppertal 1, Germany

    Klaus Pissowotzki, Kambiz Mansouri & Wolfgang Piepersberg

Authors
  1. Klaus Pissowotzki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kambiz Mansouri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wolfgang Piepersberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by H. Hennecke

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pissowotzki, K., Mansouri, K. & Piepersberg, W. Genetics of streptomycin production in Streptomyces griseus: molecular structure and putative function of genes strELMB2N . Molec. Gen. Genet. 231, 113–123 (1991). https://doi.org/10.1007/BF00293829

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00293829

Key words

Search

Navigation