The Function of 80 S Ribosomal Subunits and Effects of Some Antibiotics

Vázquez, David; Battaner, E.; Neth, R.; Heller, G; Monro, R.E.

doi:10.1101/sqb.1969.034.01.043

Cited by 65 publications

(14 citation statements)

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“…The low level of fMet-tRNA hydrolysis occurring without added tRNA may be due to tRNA that contaminates the ribosomal preparations. Sparsomycin, a potent inhibitor of reticulocyte (21) and E. coli (20) peptidyl transferase, inhibits fMet-tRNA hydrolysis in the presence Table 2 The hydrolysis of fMet-tRNA that occurs with R at peptide chain termination is somewhat similar to that occurring with tRNA and acetone (Fig. 3).…”

Section: Preparation Of F[ H]mentioning

confidence: 99%

“…Two antibiotics, sparsomycin and gougerotin, are inhibitors of the three reactions by E. coli and reticulocyte ribosomes. Both antibiotics were reported (20,21) to be inhibitors of eukaryotic and prokaryotic peptidyl transferase. Amicetin and chloramphenicol inhibit formation of fMet-ethyl ester, fMet-puromycin, and hydrolysis of fMet-tRNA by E. coli ribosomes.…”

Section: Preparation Of F[ H]mentioning

confidence: 99%

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Hydrolysis of fMet-tRNA by Peptidyl Transferase

Caskey

Beaudet

Scolnick

et al. 1971

Proc. Natl. Acad. Sci. U.S.A.

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Release factor-dependent hydrolysis of the peptidyl-tRNA ester bond requires location of the peptidyl-tRNA in the ribosomal P-site (2), NH4+ or K+ (4), and is inhibited by several antibiotics that also inhibit peptide bond formation (4-6). These characteristics are similar to those of the ribosomal enzyme peptidyl transferase. Such circumstantial information has led several investigators (4-7) to speculate on the possible involvement of peptidyl transferase in peptidyltRNA hydrolysis occurring at chain termination. Our interest in the mechanism of peptide chain termination led us to the study of peptidyl transferase. Monro and Vasquez (8) have greatly simplified study of peptidyl transferase and conducted extensive studies characterizing itsactivity. These investigators measure the ribosomal-dependent formation of fMet-puromycin from fMettRNA or 3'-terminal fragments of fMet-tRNA as an index of peptidyl transferase activity. Such reactions proceed in vitro with fMet-tRNA, K+ or NH4+, puromycin, 50S (bacterial) or 60S (eukaryotic) ribosomal subunits, and ethanol (9). Thus, peptidyl transferase activity is a function of the larger ribosomal subunit and can be studied independently When reactions contain fMet-tRNA, 50S (bacterial) ribosomal subunits, ethanol, and tRNA or its 3'-terminal oligonucleotide CpCpA, the reaction product is fMet-ethyl ester (10). Using different experimental approaches, ribosomal-dependent ester formation has been observed by Rich and his associates (7,11,12). The characteristics of both reactions indicate that transesterification is catalyzed by peptidyl transferase. This report describes the hydrolysis of ribosomal-bound fMet-tRNA by peptidyl transferase in reactions containing acetone and tRNA. The demonstration of the hydrolytic capacity of peptidyl transferase provides additional indirect evidence for its involvement in the peptide chain termination event.

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Section: Preparation Of F[ H]mentioning

confidence: 99%

Section: Preparation Of F[ H]mentioning

confidence: 99%

Hydrolysis of fMet-tRNA by Peptidyl Transferase

Caskey

Beaudet

Scolnick

et al. 1971

Proc. Natl. Acad. Sci. U.S.A.

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show abstract

“…It acts by blocking peptide-bond formation [16][17][18][19][20]. However, if the aminoacyl-tRNA in the donor site contains a free a-amino group, the inhibition cannot occur [8].…”

Section: Resultsmentioning

confidence: 99%

Aminoacyl‐tRNA binding sites in E. coli and reticulocyte ribosomes

Busiello¹,

Girolamo²

1973

FEBS Letters

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“…The peptidyl transferase centre, responsible for the catalytic activity required for the peptide bond formation step in protein synthesis, is an integral part of the larger ribosomal subunit in procaryotic [ 1 ] ns well as eucaryotic [2] cells. To investigate this Pctive centre we have developed adequate systeI;ls to study: (a) peptidyl transferase catalytic activiLy, (b) substrate interaction at the P-and A-sites of the centre and (c) interaction of a number of antibiotics.…”

Section: Introductionmentioning

confidence: 99%