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. 1973 Sep;4(3):205–213. doi: 10.1128/aac.4.3.205

Protein Synthesis in Mycobacterium tuberculosis H37Rv and the Effect of Streptomycin in Streptomycin-Susceptible and -Resistant Strains

M S Shaila 1, K P Gopinathan 1, T Ramakrishnan 1
PMCID: PMC444530  PMID: 4202339

Abstract

An efficient in vitro amino acid-incorporating system from Mycobacterium tuberculosis H37Rv was standardized. Ribonucleic acid (RNA) isolated from phage-infected M. smegmatis cells served as natural messenger RNA and directed the incorporation of 14C-amino acids into protein. The effects of various antitubercular drugs and “known inhibitors” of protein synthesis on amino acid incorporation were studied. Antibiotics like chloramphenicol and tetracycline inhibited mycobacterial protein synthesis, though they failed to prevent the growth of the organism. This failure was shown to be due to the impermeability of mycobacteria to these drugs by use of “membrane-active” agents along with the antibiotics in growth inhibition studies. Several independent streptomycin-resistant mutants of M. tuberculosis H37Rv were isolated. Streptomycin inhibited the incorporation of 14C-amino acids into proteins by whole cells of a streptomycin-susceptible strain by more than 90%, whereas very little or no inhibition was observed in either high-level or low-level streptomycin-resistant strains.

In vitro, streptomycin was an effective inhibitor of susceptible strains, whereas in streptomycin-resistant strains the concentration of streptomycin at which half-maximal inhibition was produced varied according to the resistance of whole cells, and there was a correlation between the two. In one low-level streptomycin-resistant mutant, the in vitro amino acid-incorporating system was as sensitive to various concentrations of streptomycin as the parental type, and a possible involvement of a membrane site in the development of low-level resistance was indicated. Streptomycin susceptibility and high-level resistance were shown to be ribosomal in nature.

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Selected References

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

  1. Bayer M. E., Anderson T. F. The surface structure of Escherichia coli. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1592–1599. doi: 10.1073/pnas.54.6.1592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CERIOTTI G. Determination of nucleic acids in animal tissues. J Biol Chem. 1955 May;214(1):59–70. [PubMed] [Google Scholar]
  3. Calvori C., Frontali L., Leoni L., Tecce G. Effect of rifamycin on protein synthesis. Nature. 1965 Jul 24;207(995):417–418. doi: 10.1038/207417a0. [DOI] [PubMed] [Google Scholar]
  4. ERDOS T., ULLMANN A. Effect of streptomycin on the incorporation of amino-acids labelled with carbon-14 into ribonucleic acid and protein in a cell-free system of a Mycobacterium. Nature. 1959 Feb 28;183(4661):618–619. doi: 10.1038/183618a0. [DOI] [PubMed] [Google Scholar]
  5. ERDOS T., ULLMANN A. Effect of streptomycin on the incorporation of tyrosine labelled with carbon-14 into protein of Mycobacterium cell fractions in vivo. Nature. 1960 Jan 9;185:100–101. doi: 10.1038/185100a0. [DOI] [PubMed] [Google Scholar]
  6. FLAKS J. G., COX E. C., WHITE J. R. Inhibition of polypeptide synthesis by streptomycin. Biochem Biophys Res Commun. 1962 May 11;7:385–389. doi: 10.1016/0006-291x(62)90320-0. [DOI] [PubMed] [Google Scholar]
  7. FLAKS J. G., COX E. C., WITTING M. L., WHITE J. R. Polypeptide synthesis with ribosomes from streptomycin-resistant and dependent E. coli. Biochem Biophys Res Commun. 1962 May 11;7:390–393. doi: 10.1016/0006-291x(62)90321-2. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lucas-Lenard J. Protein biosynthesis. Annu Rev Biochem. 1971;40:409–448. doi: 10.1146/annurev.bi.40.070171.002205. [DOI] [PubMed] [Google Scholar]
  10. Morgan A. R., Wells R. D., Khorana H. G. Studies on polynucleotides, lix. Further codon assignments from amino Acid incorporations directed by ribopolynucleotides containing repeating trinucleotide sequences. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1899–1906. doi: 10.1073/pnas.56.6.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pestka S. Inhibitors of ribosome functions. Annu Rev Microbiol. 1971;25:487–562. doi: 10.1146/annurev.mi.25.100171.002415. [DOI] [PubMed] [Google Scholar]
  12. Raj C. V., Ramakrishnan T. Transduction in Mycobacterium smegmatis. Nature. 1970 Oct 17;228(5268):280–281. doi: 10.1038/228280b0. [DOI] [PubMed] [Google Scholar]
  13. Ramakrishnan T., Murthy P. S., Gopinathan K. P. Intermediary metabolism of mycobacteria. Bacteriol Rev. 1972 Mar;36(1):65–108. doi: 10.1128/br.36.1.65-108.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rieber M., Imaeda T. Effect of isoniazid on mycobacterial polyphenylalanine synthesis. Biochim Biophys Acta. 1969 Jul 22;186(1):173–177. doi: 10.1016/0005-2787(69)90500-0. [DOI] [PubMed] [Google Scholar]
  15. SPEYER J. F., LENGYEL P., BASILIO C. Ribosomal localization of streptomycin sensitivity. Proc Natl Acad Sci U S A. 1962 Apr 15;48:684–686. doi: 10.1073/pnas.48.4.684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Salser W., Gesteland R. F., Bolle A. In vitro synthesis of bacteriophage lysozyme. Nature. 1967 Aug 5;215(5101):588–591. doi: 10.1038/215588a0. [DOI] [PubMed] [Google Scholar]
  17. Sriprakash K. S., Ramakrishnan T. Isoniazid-resistant mutants of Mycobacterium tuberculosis H37RV: uptake of isoniazid and the properties of NADase inhibitor. J Gen Microbiol. 1970 Jan;60(1):125–132. doi: 10.1099/00221287-60-1-125. [DOI] [PubMed] [Google Scholar]
  18. Stuart J. J., Ravin A. W. Levels of resistance in ribosomes from genetically linked, streptomycin-resistant mutants of pneumococcus. J Gen Microbiol. 1968 May;51(3):411–424. doi: 10.1099/00221287-51-3-411. [DOI] [PubMed] [Google Scholar]
  19. Trnka L., Smith D. W. Proteosynthetic activity of isolated ribosomes of Mycobacteria and its alteration by rifampicin and related tuberculostatic drugs. Antibiot Chemother. 1970;16:369–379. doi: 10.1159/000386840. [DOI] [PubMed] [Google Scholar]
  20. van Dijk-Salkinoja M. S., Planta R. J. Rate of ribosome production in Bacillus licheniformis. J Bacteriol. 1971 Jan;105(1):20–27. doi: 10.1128/jb.105.1.20-27.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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