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. 1988 Jan;7(1):277–282. doi: 10.1002/j.1460-2075.1988.tb02810.x

Resistance to macrolides, lincosamides and streptogramin type B antibiotics due to a mutation in an rRNA operon of Streptomyces ambofaciens.

J L Pernodet 1, F Boccard 1, M T Alegre 1, M H Blondelet-Rouault 1, M Guérineau 1
PMCID: PMC454269  PMID: 2834204

Abstract

Streptomyces ambofaciens produces spiramycin, a macrolide antibiotic and expresses an inducible resistance to macrolides, lincosamides and streptogramin B antibiotics (MLS). From a mutant of S.ambofaciens exhibiting a constitutive MLS resistance phenotype a resistance determinant was cloned on a low copy number vector (pIJ61) through its expression in Streptomyces lividans. Further characterization has shown that this determinant corresponded to a mutant rRNA operon with a mutation in the 23S rRNA gene. In different organisms, mutations leading to MLS resistance have been located at a position corresponding to the adenine 2058 of Escherichia coli 23S rRNA. In the 23S rRNA from S.ambofaciens a similar position for the mutation has been postulated and DNA sequencing of this region has shown an adenine to guanine transition at a position corresponding to 2058. S.ambofaciens possesses four rRNA operons which we have cloned. In Streptomyces, contrary to other bacteria, a mutation in one among several rRNA operons confers a selectable MLS resistance phenotype. Possible reasons for this difference are discussed.

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Selected References

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  1. Ahmed A. Mechanism of inhibition of protein synthesis by spiramycin. Biochim Biophys Acta. 1968 Aug 23;166(1):205–217. doi: 10.1016/0005-2787(68)90504-2. [DOI] [PubMed] [Google Scholar]
  2. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birmingham V. A., Cox K. L., Larson J. L., Fishman S. E., Hershberger C. L., Seno E. T. Cloning and expression of a tylosin resistance gene from a tylosin-producing strain of Streptomyces fradiae. Mol Gen Genet. 1986 Sep;204(3):532–539. doi: 10.1007/BF00331036. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  6. Cundliffe E., Thompson J. The mode of action of nosiheptide (multhiomycin) and the mechanism of resistance in the producing organism. J Gen Microbiol. 1981 Sep;126(1):185–192. doi: 10.1099/00221287-126-1-185. [DOI] [PubMed] [Google Scholar]
  7. Ettayebi M., Prasad S. M., Morgan E. A. Chloramphenicol-erythromycin resistance mutations in a 23S rRNA gene of Escherichia coli. J Bacteriol. 1985 May;162(2):551–557. doi: 10.1128/jb.162.2.551-557.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fujisawa Y., Weisblum B. A family of r-determinants in Streptomyces spp. that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibiotics. J Bacteriol. 1981 May;146(2):621–631. doi: 10.1128/jb.146.2.621-631.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham M. Y., Weisblum B. 23S ribosomal ribonucleic acid of macrolide-producing streptomycetes contains methylated adenine. J Bacteriol. 1979 Mar;137(3):1464–1467. doi: 10.1128/jb.137.3.1464-1467.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HICKEY R. J., TRESNER H. D. A cobalt-containing medium for sporulation of Streptomyces species. J Bacteriol. 1952 Dec;64(6):891–892. doi: 10.1128/jb.64.6.891-892.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  12. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  13. Hopwood D. A., Hintermann G., Kieser T., Wright H. M. Integrated DNA sequences in three streptomycetes form related autonomous plasmids after transfer to Streptomyces lividans. Plasmid. 1984 Jan;11(1):1–16. doi: 10.1016/0147-619x(84)90002-7. [DOI] [PubMed] [Google Scholar]
  14. Hopwood D. A., Kieser T., Wright H. M., Bibb M. J. Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol. 1983 Jul;129(7):2257–2269. doi: 10.1099/00221287-129-7-2257. [DOI] [PubMed] [Google Scholar]
  15. Lydiate D. J., Malpartida F., Hopwood D. A. The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors. Gene. 1985;35(3):223–235. doi: 10.1016/0378-1119(85)90001-0. [DOI] [PubMed] [Google Scholar]
  16. Maccecchini M. L., Rudin Y., Blobel G., Schatz G. Import of proteins into mitochondria: precursor forms of the extramitochondrially made F1-ATPase subunits in yeast. Proc Natl Acad Sci U S A. 1979 Jan;76(1):343–347. doi: 10.1073/pnas.76.1.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  19. Noller H. F. Structure of ribosomal RNA. Annu Rev Biochem. 1984;53:119–162. doi: 10.1146/annurev.bi.53.070184.001003. [DOI] [PubMed] [Google Scholar]
  20. Omura S., Nakagawa A., Otani M., Hati T., Ogura H. Structure of the spiramycins (foromacidines) and their relationship with the leucomycins and carbomycins (magnamycins). J Am Chem Soc. 1969 Jun 4;91(12):3401–3404. doi: 10.1021/ja01040a069. [DOI] [PubMed] [Google Scholar]
  21. Richter K., Ammerer G., Hartter E., Ruis H. The effect of delta-aminolevulinate on catalase T-messenger RNA levels in delta-aminolevulinate synthase-defective mutants of Saccharomyces cerevisiae. J Biol Chem. 1980 Sep 10;255(17):8019–8022. [PubMed] [Google Scholar]
  22. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Sigmund C. D., Ettayebi M., Morgan E. A. Antibiotic resistance mutations in 16S and 23S ribosomal RNA genes of Escherichia coli. Nucleic Acids Res. 1984 Jun 11;12(11):4653–4663. doi: 10.1093/nar/12.11.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sigmund C. D., Morgan E. A. Erythromycin resistance due to a mutation in a ribosomal RNA operon of Escherichia coli. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5602–5606. doi: 10.1073/pnas.79.18.5602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Sor F., Fukuhara H. Identification of two erythromycin resistance mutations in the mitochondrial gene coding for the large ribosomal RNA in yeast. Nucleic Acids Res. 1982 Nov 11;10(21):6571–6577. doi: 10.1093/nar/10.21.6571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thompson C. J., Kieser T., Ward J. M., Hopwood D. A. Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene. 1982 Nov;20(1):51–62. doi: 10.1016/0378-1119(82)90086-5. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Thompson C. J., Ward J. M., Hopwood D. A. Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol. 1982 Aug;151(2):668–677. doi: 10.1128/jb.151.2.668-677.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  34. Weisblum B., Siddhikol C., Lai C. J., Demohn V. Erythromycin-inducible resistance in Staphylococcus aureus: requirements for induction. J Bacteriol. 1971 Jun;106(3):835–847. doi: 10.1128/jb.106.3.835-847.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wittmann H. G., Stöffler G., Apirion D., Rosen L., Tanaka K., Tamaki M., Takata R., Dekio S., Otaka E. Biochemical and genetic studies on two different types of erythromycin resistant mutants of Escherichia coli with altered ribosomal proteins. Mol Gen Genet. 1973 Dec 20;127(2):175–189. doi: 10.1007/BF00333665. [DOI] [PubMed] [Google Scholar]
  36. 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]

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