Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Dec;174(23):7868–7872. doi: 10.1128/jb.174.23.7868-7872.1992

Cloning and characterization of an aminoglycoside resistance determinant from Micromonospora zionensis.

M Kojic 1, L Topisirovic 1, B Vasiljevic 1
PMCID: PMC207509  PMID: 1447159

Abstract

The sisomicin-gentamicin resistance methylase (sgm) gene was isolated from Micromonospora zionensis and cloned in Streptomyces lividans. The sgm gene was expressed in Micromonospora melanosporea, where its own promoter was active, and also in Escherichia coli under the control of the lacZ promoter. The complete nucleotide sequence of 1,122 bp and a transcription start point were determined. The sequence contains an open reading frame that encodes a polypeptide of 274 amino acids. The methylation of 30S ribosomal subunits by Sgm methylase accounts adequately for all known resistance characteristics of M. zionensis, but expression of high-level resistance to hygromycin B is background dependent. A comparison of the amino acid sequence of the predicted Sgm protein with the deduced amino acid sequences for the 16S rRNA methylases showed extensive similarity of Grm and significant similarity to KgmB but not to KamB methylase.

Full text

PDF
7868

Images in this article

7871Image
on p.7871

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baum E. Z., Love S. F., Rothstein D. M. Temporally regulated tandem promoters in Micromonospora echinospora. J Bacteriol. 1988 Jan;170(1):71–77. doi: 10.1128/jb.170.1.71-77.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beauclerk A. A., Cundliffe E. Sites of action of two ribosomal RNA methylases responsible for resistance to aminoglycosides. J Mol Biol. 1987 Feb 20;193(4):661–671. doi: 10.1016/0022-2836(87)90349-4. [DOI] [PubMed] [Google Scholar]
  3. Cundliffe E. How antibiotic-producing organisms avoid suicide. Annu Rev Microbiol. 1989;43:207–233. doi: 10.1146/annurev.mi.43.100189.001231. [DOI] [PubMed] [Google Scholar]
  4. Distler J., Braun C., Ebert A., Piepersberg W. Gene cluster for streptomycin biosynthesis in Streptomyces griseus: analysis of a central region including the major resistance gene. Mol Gen Genet. 1987 Jun;208(1-2):204–210. doi: 10.1007/BF00330443. [DOI] [PubMed] [Google Scholar]
  5. Gough J. A., Murray N. E. Sequence diversity among related genes for recognition of specific targets in DNA molecules. J Mol Biol. 1983 May 5;166(1):1–19. doi: 10.1016/s0022-2836(83)80047-3. [DOI] [PubMed] [Google Scholar]
  6. Haydock S. F., Dowson J. A., Dhillon N., Roberts G. A., Cortes J., Leadlay P. F. Cloning and sequence analysis of genes involved in erythromycin biosynthesis in Saccharopolyspora erythraea: sequence similarities between EryG and a family of S-adenosylmethionine-dependent methyltransferases. Mol Gen Genet. 1991 Nov;230(1-2):120–128. doi: 10.1007/BF00290659. [DOI] [PubMed] [Google Scholar]
  7. Holmes D. J., Cundliffe E. Analysis of a ribosomal RNA methylase gene from Streptomyces tenebrarius which confers resistance to gentamicin. Mol Gen Genet. 1991 Oct;229(2):229–237. doi: 10.1007/BF00272160. [DOI] [PubMed] [Google Scholar]
  8. Holmes D. J., Drocourt D., Tiraby G., Cundliffe E. Cloning of an aminoglycoside-resistance-encoding gene, kamC, from Saccharopolyspora hirsuta: comparison with kamB from Streptomyces tenebrarius. Gene. 1991 Jun 15;102(1):19–26. doi: 10.1016/0378-1119(91)90532-g. [DOI] [PubMed] [Google Scholar]
  9. Katz E., Thompson C. J., Hopwood D. A. Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol. 1983 Sep;129(9):2703–2714. doi: 10.1099/00221287-129-9-2703. [DOI] [PubMed] [Google Scholar]
  10. Kelemen G. H., Cundliffe E., Financsek I. Cloning and characterization of gentamicin-resistance genes from Micromonospora purpurea and Micromonospora rosea. Gene. 1991 Feb 1;98(1):53–60. doi: 10.1016/0378-1119(91)90103-i. [DOI] [PubMed] [Google Scholar]
  11. Moazed D., Noller H. F. Interaction of antibiotics with functional sites in 16S ribosomal RNA. Nature. 1987 Jun 4;327(6121):389–394. doi: 10.1038/327389a0. [DOI] [PubMed] [Google Scholar]
  12. Piendl W., Böck A., Cundliffe E. Involvement of 16S ribosomal RNA in resistance of the aminoglycoside-producers Streptomyces tenjimariensis, Streptomyces tenebrarius and Micromonospora purpurea. Mol Gen Genet. 1984;197(1):24–29. doi: 10.1007/BF00327918. [DOI] [PubMed] [Google Scholar]
  13. Piendl W., Böck A. Ribosomal resistance in the gentamicin producer organism Micromonospora purpurea. Antimicrob Agents Chemother. 1982 Aug;22(2):231–236. doi: 10.1128/aac.22.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Salauze D., Davies J. Isolation and characterisation of an aminoglycoside phosphotransferase from neomycin-producing Micromonospora chalcea; comparison with that of Streptomyces fradiae and other producers of 4,6-disubstituted 2-deoxystreptamine antibiotics. J Antibiot (Tokyo) 1991 Dec;44(12):1432–1443. doi: 10.7164/antibiotics.44.1432. [DOI] [PubMed] [Google Scholar]
  15. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Skeggs P. A., Thompson J., Cundliffe E. Methylation of 16S ribosomal RNA and resistance to aminoglycoside antibiotics in clones of Streptomyces lividans carrying DNA from Streptomyces tenjimariensis. Mol Gen Genet. 1985;200(3):415–421. doi: 10.1007/BF00425725. [DOI] [PubMed] [Google Scholar]
  17. Skinner R., Cundliffe E., Schmidt F. J. Site of action of a ribosomal RNA methylase responsible for resistance to erythromycin and other antibiotics. J Biol Chem. 1983 Oct 25;258(20):12702–12706. [PubMed] [Google Scholar]
  18. Spangler E. A., Blackburn E. H. The nucleotide sequence of the 17S ribosomal RNA gene of Tetrahymena thermophila and the identification of point mutations resulting in resistance to the antibiotics paromomycin and hygromycin. J Biol Chem. 1985 May 25;260(10):6334–6340. [PubMed] [Google Scholar]
  19. Thompson C. J., Skinner R. H., Thompson J., Ward J. M., Hopwood D. A., Cundliffe E. Biochemical characterization of resistance determinants cloned from antibiotic-producing streptomycetes. J Bacteriol. 1982 Aug;151(2):678–685. doi: 10.1128/jb.151.2.678-685.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thompson J., Skeggs P. A., Cundliffe E. Methylation of 16S ribosomal RNA and resistance to the aminoglycoside antibiotics gentamicin and kanamycin determined by DNA from the gentamicin-producer, Micromonospora purpurea. Mol Gen Genet. 1985;201(2):168–173. doi: 10.1007/BF00425655. [DOI] [PubMed] [Google Scholar]
  21. Wagman G. H., Weinstein M. J. Antibiotic from Micromonospora. Annu Rev Microbiol. 1980;34:537–557. doi: 10.1146/annurev.mi.34.100180.002541. [DOI] [PubMed] [Google Scholar]
  22. Westpheling J., Ranes M., Losick R. RNA polymerase heterogeneity in Streptomyces coelicolor. Nature. 1985 Jan 3;313(5997):22–27. doi: 10.1038/313022a0. [DOI] [PubMed] [Google Scholar]
  23. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES