Summary
An in vitro polypeptide synthesis system was set up for three methanogenic bacteria, Methanococcus vannielii, Methanobacterium formicicum and Methanosarcina barkeri, and the effect of classical 70S and 80S protein synthesis inhibitors studied. The following results were obtained: (i) The activity of ribosomes from all three methanogens was unaffected by a number of 70S inhibitors such as tetracycline, chloramphenicol, streptomycin, tiamulin and, probably, erythromycin as well; (ii) However, the ribosomes were sensitive to thiostrepton, virginiamycin and, to varying degrees, to those aminoglycosides containing a 2-deoxystreptamine moiety. Among the aminoglycosides examined, streptomycin induced no translational misreading. The compounds containing 2-deoxystreptamine stimulated misreading, albeit only at high concentrations (neomycin being an exception); (iii) Ribosomes from all three organisms were insensitive to the 80S inhibitors cycloheximide and ricin, but those from Methanobacterium formicicum were highly sensitive to anisomycin and moderately sensitive to verrucarin. The results support those of in vivo studies and provide conclusive evidence that archaebacterial ribosomes despite being 70S ribosomes lack binding sites for many classical eubacterial ribosome inhibitors. At the same time they possess sites for others, as well as for some inhibitors of 80S ribosomes.
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References
Barbacid M, Vazquez D (1975) Ribosome changes during translation. J Mol Biol 93:449–463
Böck A, Petzet A, Piepersberg W (1979) Ribosomal ambiguity (ram) mutations facilitate dihydrosteptomycin binding to ribosomes. FEBS Lett 104:317–321
Bryan LE, Van den Elzen HM (1977) Effect of membrane energy mutations and cations on streptomycin and gentamicin accumulation by bacteria: a model for entry of steptomycin and gentamicin in susceptible and resistant bacteria. Antimicrob Agents Chemother 12:163–177
Chang FN, Flaks JG (1972) The binding of dihydrostreptomycin to E. coli ribosomes. Characteristics and equilibrium of the reaction. Antimicrob Agents Chemother 2:294–307
Cocito C (1979) Antibiotics of the virginiamycin family, inhibitors which contain synergistic components. Microbiol Rev 43:145–198
Cundliffe E (1979) Thiostrepton and related antibiotics. In: Hahn FE (ed) Antibiotics V-2. Mechanism of action of antieukaryotic and antiviral compounds. Springer, Berlin Heidelberg New York, pp 329–343
Davies J, Davis BD (1968) Misreading of ribonucleic acid code words induced by aminoglycoside antibiotics. The effect of drug concentration. J Biol Chem 243:3312–3316
Douglas C, Achatz F, Böck A (1980) Electrophoretic characterisation of ribosomal proteins from methanogenic bacteria. Zbl Bakt Hyg I Abt Orig C 1:1–11
Ehrenstein G von (1967) Isolation of sRNA from intact Escherichia coli cells. In: Grossman L, Moldave K (eds) Methods Enzymol, vol XII, p 588–596
Fox GE, Stackebrandt E, Hespell RB, Gibson J, Maniloff J, Dryer TA, Wolfe RS, Balch WE, Tanner RS, Magrum RJ, Zablen LB, Blakemore R, Grupta R, Bonen L, Lewis JB, Stahl AD, Luehrsen KR, Chen KN, Woese CR (1980) The phylogeny of prokaryotes. Science 209:457–463
Greco M, Montanaro L, Novello F, Saccone C, Sperti S, Stirpe F (1974) Inhibition of protein synthesis by ricin: experiments with rat liver mitochondria and nuclei and with ribosomes from Escherichia coli. Biochem J 142:695–697
Grisé-Miron L, Noreau J, Melançon P, Brakier-Gingras L (1981) Comparism of the misreading induced by streptomycin and neomycin. Biochim Biophys Acta 656:103–110
Hilpert R, Winter J, Hammes W, Kandler O (1981) The sensitivity of archaebacteria to antibiotics. Zbl Bakt Hyg I Abt Orig C2:11–20
Högenauer G (1979) Tiamulin and Pleuromutilin. In: Hahn FE (ed) Antibiotics V-2. Mechanism of action of antieukaryotic and antiviral compounds. Springer, Berlin Heidelberg New York, pp 344–360
Kaji A, Ryoji M (1979) Tetracycline. In: Hahn FE (ed) Antibiotics V-2. Mechanism of action of antieukaryotic and antiviral compounds. Springer, Berlin Heidelberg New York, pp 304–328
Kućan Ž, Lipmann F (1964) Differences in chloramphenicol sensitivity of cell-free amino acid polymerization systems. J Biol Chem 239:516–520
McKellar RC, Sprott GD (1979) Solubilization and properties of a particulate hydrogenase from Methanobacterium strain G2R. J Bacteriol 139:231–238
Pecher T, Böck A (1981) In vivo susceptibility of halophilic and methanogenic organisms to protein synthesis inhibitors. FEMS Microbiol Lett 10:295–297
Rosset R, Gorini L (1969) A ribosomal ambiguity mutation. J Mol Biol 39:95–112
Schindler D (1974) Two classes of inhibitors of peptidyl transferase activity in eukaryotes. Nature 249:38–41
Schmid G, Pecher T, Böck A (1982) Properties of the translational apparatus of archaebacteria. Zbl Bakt Hyg I Abt. Orig C 3:209–217
Stöcklein W, Piepersberg W (1980) Altered ribosomal protein L29 in a cycloheximide-resistant strain of Saccharomyces cerevisiae. Curr Genetics 1:177–183
Tai PC, Davis BD (1979) Triphasic concentration effects of gentamicin on activity and misreading in protein synthesis. Biochemistry 18:193–198
Wilhelm JM, Pettitt SE, Jessop JJ (1978) Aminoglycoside antibiotics and eukaryotic protein synthesis: Structure-function relationships in the stimulation of misreading with a wheat embryo system. Biochemistry 17:1143–1149
Zierhut G, Piepersberg W, Böck A (1979) Comparative analysis of the effect of aminoglycosides on bacterial protein synthesis in vitro. Eur J Biochem 98:577–583
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Elhardt, D., Böck, A. An in vitro polypeptide synthesizing system from methanogenic bacteria: Sensitivity to antibiotics. Molec Gen Genet 188, 128–134 (1982). https://doi.org/10.1007/BF00333006
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DOI: https://doi.org/10.1007/BF00333006