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Streptomycin-sensitivity in Streptomyces glaucescens is due to deletions comprising the structural gene coding for a specific phosphotransferase

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Summary

The wild type strain of Streptomyces glaucescens produces hydroxystreptomycin and has a natural resistance towards the streptomycin group aminoglycoside antibiotics. The inherent resistance is a genetically unstable character and mutant strains sensitive to streptomycins arise spontaneously at unusually high frequencies. The gene conferring streptomycin resistance was cloned and characterised as a streptomycin specific phosphotransferase. Hybridisation experiments show that the mutational event leading to sensitivity is due to large deletions, most likely on the chromosome, comprehending the structural gene coding for a streptomycin phosphotransferase and its flanking regions. Interspecific expression of the S. glaucescens phosphotransferase was found in Streptomyces lividans as well as in Escherichia coli.

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Abbreviations

bp:

base pairs

EDTA:

ethylenediaminetetraacetic acid

kb:

kilobases'

TES:

n-tris(hydroxymethyl) methyl-2-aminoethane sulfonic acid

References

  • Baumann R (1973) Genetik von Streptomyces glaucescens. Ph D thesis, Eidgenössische Technische Hochschule Zürich, no 4968

  • Baumann R, Hütter R, Hopwood DA (1974) Genetics of Streptomyces glaucescens and regulation of melanin production. J Gen Microbiol 81:463–474

    Google Scholar 

  • Bibb MJ, Ward JM, Hopwood DA (1978) Tranformation of plasmid DNA into Streptomyces at high frequency. Nature London 274:398–400

    Google Scholar 

  • Bolivar F (1978) Construction and characterisation of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique EcoRI sites for selection of EcoRI-generated recombinant DNA molecules. Gene 4:121–136

    Google Scholar 

  • Chater KF, Hopwood DA, Kieser T, Thompson CJ (1982) Gene cloning in Streptomyces. Curr Topics Microbiol Immunol 97:69–95

    Google Scholar 

  • Crameri R (1981) Genetik von Streptomyces glaucescens unter besonderer Berücksichtigung chromosomaler Instabilitäten. Ph D thesis, Eidgenössische Technische Hochschule Zürich, no 6884

  • Crameri R, Kieser T, Ono H, Sanchez J, Hütter R (1983) Chromosomal instability in Streptomyces glaucescens: mapping of streptomycin-sensitive mutants. J Gen Microbiol 129:519–527

    Google Scholar 

  • Davies J, Houk C, Yagisawa M, White TJ (1979) Occurrence and functions of aminoglycoside-modifying enzymes. In: Sebek OK, Laskin AI (eds) Genetics of industrial microorganisms. American Soceity for Microbiology, Washington DC, p 166

    Google Scholar 

  • Davies J, Smith DI (1978) Plasmid-determined resistance to antimicrobial agents. Annu Rev Microbiol 32:469–518

    Google Scholar 

  • Fangman WL (1978) Separation of very large DNA molecules by gel electrophoresis. Nucl Acid Res 5:653–665

    Google Scholar 

  • Freeman RF, Hopwood DA (1978) Unstable naturally occurring resistance to antibiotics in Streptomyces. J Gen Microbiol 106:377–381

    Google Scholar 

  • Hattman S, Brooks JE, Masurekar M (1978) Sequence specificity of the P1 modification methylase (M. EcoP1) and the DNA methylase (M. Eco dam) controlled by Escherichia coli dam gene. J Mol Biol 126:367–380

    Google Scholar 

  • Hintermann G, Crameri R, Kieser T, Hütter R (1981) Restriction analysis of the Streptomyces glaucescens genome by agarose gel electrophoresis. Arch Microbiol 130:218–222

    Google Scholar 

  • Hopwood DA, Sermonti G (1962) The genetics of Streptomyces coelicolor. Adv Genet 11:273–342

    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 

  • Horinouchi S, Uozumi T, Beppu T (1980) Cloning of Streptomyces DNA into Escherichia coli: absence of heterospecific gene expression of Streptomyces genes in E. coli. Agric Biol Chem 44:367–381

    Google Scholar 

  • Hütter R (1967) Systematik der Streptomyceten. Karger Verlag, Basel

    Google Scholar 

  • Hütter R, Kieser T, Crameri R, Hintermann G (1981) Chromosomal instability in Streptomyces glaucescens. Zbl Bakt Suppl 11:551–559

    Google Scholar 

  • Kieser T (1979) Untersuchungen von genetisch instabilen Eigenschaften von Streptomyces glaucescens. Ph D thesis, Eidgenössische Technische Hochschule Zürich, no 6473

  • 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 

  • Low B (1968) Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci USA 60:160–167

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Miller AL, Walker JB (1969) Enzymatic phosphorylation of streptomycin by extracts of streptomycin-producing strains of Streptomyces. J Bacteriol 99:401–405

    Google Scholar 

  • Okanishi M, Suzuki K, Umezawa H (1974) Formation and reversion of streptomycete protoplasts. Cultural conditions and morphological study. J Gen Microbiol 80:389–400

    Google Scholar 

  • Ono H, Crameri R, Hintermann G, Hütter R (1983) Hydroxystreptomycin production and resistance in Streptomyces glaucescens. J Gen Microbiol 129:529–537

    Google Scholar 

  • Pridham TG, Anderson P, Foley C, Lindensfelder LA, Benedict RG (1956/57) A selection of media for maintenance and taxonomic study of Streptomyces. Antibiotics Ann 947-953

  • Robbins PW, Wirth DF, Hering C (1981) Expression of the Streptomyces enzyme endoglycosidase H in Escherichia coli. J Biol Chem 256:10640–10644

    Google Scholar 

  • Rodgers WH, Springer W, Young FE (1982) Cloning and expression of a Streptomyces fradiae neomycin resistance gene in Escherichia coli. Gene 18:133–141

    Google Scholar 

  • Schrempf H, Bujard H, Hopwood DA, Goebel W (1975) Isolation of covalently closed circular deoxyribonucleic acid from Streptomyces coelicolor A3(2). J Bacteriol 121:416–421

    Google Scholar 

  • Schrempf H (1983) Deletion and amplification of DNA sequences in melanin-negative variants of Streptomyces reticuli. Mol Gen Genet 189:501–505

    Google Scholar 

  • Schupp T, Toupet C, Stalhammer-Carlemalm M, Meyer J (1983) Expression of a neomycin phosphotransferase gene from Streptomyces fradiae in Escherichia coli after interplasmatic recombination. Mol Gen Genet 189:27–33

    Google Scholar 

  • Suter MA (1978) Isolierung und Charakterisierung von Melaninnegativen Mutanten aus Streptomyces glaucescens. Ph D thesis, Eidgenössische Technische Hochschule Zürich, No 6276

  • Suter MA, Hütter R, Leisinger T (1978a) Mutants of Streptomyces glaucescens affected in the production of extracellular enzymes. In: Freerksen E, Tarnok I, Thumin IH (eds) Genetics of the actinomycetales. Gustav Fischer Verlag, Stuttgart-New York, p 61

    Google Scholar 

  • Suter MA, Hütter R, Leisinger T (1978b) Induction of albino mutants in Streptomyces glaucescens. Experientia 34:1669

    Google Scholar 

  • Tabak HF, Hecht NB, Menke HH, Hollenberg CP (1979) The gene for the small ribosomal RNA on yeast mitochondrial DNA: physical map, direction of transcription and absence of an intervening sequence. Curr Genet 1:33–43

    Google Scholar 

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

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1982a) Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol 151:668–677

    Google Scholar 

  • Walker JB (1975) ATP: streptomycin 6-phosphotransferase. Methods Enzymol 43:628–632

    Google Scholar 

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Communicated by W. Arber

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Hintermann, G., Crameri, R., Vögtli, M. et al. Streptomycin-sensitivity in Streptomyces glaucescens is due to deletions comprising the structural gene coding for a specific phosphotransferase. Molec. Gen. Genet. 196, 513–520 (1984). https://doi.org/10.1007/BF00436201

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