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. 1980 Sep;77(9):5492–5496. doi: 10.1073/pnas.77.9.5492

Lasting damage to bacterial ribosomes by reversibly bound virginiamycin M.

R Parfait, C Cocito
PMCID: PMC350087  PMID: 6776538

Abstract

The M and S components of virginiamycin (VM and VS) inhibit protein synthesis in bacteria--reversibly when a single component is present and irreversibly when both are present. In cell-free systems, each factor binds to the large ribosomal subunit, and the affinity of ribosomes for VS is enhanced in the presence of VM. The present work shows that the action of VM (a 500-dalton modified depsipeptide) in vivo and in vitro persists upon its removal. The in vivo demonstration is based on the loss of viability of uninfected bacteria, and on the irreversible inactivation of virus-infected cells, that are caused by a sequential incubation with VM and VS (the inhibitory action of either component alone is reversible). In vitro, the binding of labeled VM to ribosomes, followed by its detachment, yields particles unable to perform poly(U)-directed polyphenylalanine synthesis. Also, the association constant for the binding of VS to these particles is equal to that of particles incubated with a mixture of VM and VS. Our findings indicate that VM action is catalytic rather than stoichiometric, and suggest the occurrence of two states of the large ribosomal subunit, a situation leading to a complex equilibrium with multiple transitional steps in the presence of virginiamycin.

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Selected References

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

  1. Boon T. Inactivation of ribosomes in vitro by colicin E 3 . Proc Natl Acad Sci U S A. 1971 Oct;68(10):2421–2425. doi: 10.1073/pnas.68.10.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowman C. M., Dahlberg J. E., Ikemura T., Konisky J., Nomura M. Specific inactivation of 16S ribosomal RNA induced by colicin E3 in vivo. Proc Natl Acad Sci U S A. 1971 May;68(5):964–968. doi: 10.1073/pnas.68.5.964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowman C. M., Sidikaro J., Nomura M. Specific inactivation of ribosomes by colicin E3 in vitro and mechanism of immunity in colicinogenic cells. Nat New Biol. 1971 Dec 1;234(48):133–137. doi: 10.1038/newbio234133a0. [DOI] [PubMed] [Google Scholar]
  4. Cocito C. Antibiotics of the virginiamycin family, inhibitors which contain synergistic components. Microbiol Rev. 1979 Jun;43(2):145–192. doi: 10.1128/mr.43.2.145-192.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cocito C., Di Giambattista M. The in vitro binding of virginiamycin M to bacteria ribosomes and ribosomal subunits. Mol Gen Genet. 1978 Oct 25;166(1):53–59. doi: 10.1007/BF00379729. [DOI] [PubMed] [Google Scholar]
  6. Cocito C., Kaji A. Virginiamycin M, a specific inhibitor of the acceptor site of ribosomes. Biochimie. 1971;53(6):763–770. doi: 10.1016/s0300-9084(71)80117-7. [DOI] [PubMed] [Google Scholar]
  7. Cocito C. Metabolism of macromolecules in bacteria treated with virginiamycin. J Gen Microbiol. 1969 Aug;57(2):179–194. doi: 10.1099/00221287-57-2-179. [DOI] [PubMed] [Google Scholar]
  8. Cocito C. Origin and metabolic properties of the RNA species formed during the replication cycle of virus 2C. J Virol. 1974 Dec;14(6):1482–1493. doi: 10.1128/jvi.14.6.1482-1493.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cocito C. Pressure dissociation of bacterial ribosomes and reassociation of ribosomal subunits. Mol Gen Genet. 1978 Jun 1;162(1):43–50. doi: 10.1007/BF00333849. [DOI] [PubMed] [Google Scholar]
  10. Cocito C., Voorma H. O., Bosch L. Interference of virginiamycin M with the initiation and the elongation of peptide chains in cell-free systems. Biochim Biophys Acta. 1974 Mar 27;340(3):285–298. doi: 10.1016/0005-2787(74)90274-3. [DOI] [PubMed] [Google Scholar]
  11. Contreras A., Vázquez D. Synergistic interaction of the streptogramins with the ribosome. Eur J Biochem. 1977 Apr 15;74(3):549–551. doi: 10.1111/j.1432-1033.1977.tb11423.x. [DOI] [PubMed] [Google Scholar]
  12. Ennis H. L. Binding of the antibiotic vernamycin in Balpha to Escherichia coli ribosomes. Arch Biochem Biophys. 1974 Feb;160(2):394–401. doi: 10.1016/0003-9861(74)90413-5. [DOI] [PubMed] [Google Scholar]
  13. Ennis H. L. Inhibition of protein synthesis by polypeptide antibiotics. 3. Ribosomal site of inhibition. Mol Pharmacol. 1966 Sep;2(5):444–453. [PubMed] [Google Scholar]
  14. Ennis H. L. Inhibition of protein synthesis by polypeptide antibiotics.. II. In vitro protein synthesis. J Bacteriol. 1965 Oct;90(4):1109–1119. doi: 10.1128/jb.90.4.1109-1119.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ennis H. L. Interaction of vernamycin A with Escherichia coli ribosomes. Biochemistry. 1971 Mar 30;10(7):1265–1270. doi: 10.1021/bi00783a025. [DOI] [PubMed] [Google Scholar]
  16. Parfait R., de Béthune M. P., Cocito C. A spectrofluorimetric study of the interaction between virginiamycin S and bacterial ribosomes. Mol Gen Genet. 1978 Oct 25;166(1):45–51. doi: 10.1007/BF00379728. [DOI] [PubMed] [Google Scholar]
  17. de Bethune M. P., Nierhaus K. H. Characterisation of the binding of virginiamycin S to Escherichia coli ribosomes. Eur J Biochem. 1978 May;86(1):187–191. doi: 10.1111/j.1432-1033.1978.tb12298.x. [DOI] [PubMed] [Google Scholar]

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