Initiation of protein synthesis,I. Effect of formylation of methionyl-tRNA on codon recognition.

Leder, Philip; Bursztyn, Hela

doi:10.1073/pnas.56.5.1579

Cited by 34 publications

(14 citation statements)

References 14 publications

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“…When EF-G and GTP were added subsequent to irradiation the crosslinked A c [~H ] P~~-~R N A -poly(U) complex was translocated to the P-site. Now 67 of the radioactivity was released from the Ac[~H]-Phe-tRNA -poly( U) complex by incubation with puromycin.The efficiency of translocation corresponds to that observed in a parallel nonirradiated control using the standard puromycin assay of Leder and Bursztyn [31] indicating that photo-crosslinking does not interfere with translocation. from T. utilis was prepared and its crosslinking was compared with that of S. carlshergensis used in the previous experiments.…”

Section: Translocation Of the Crosslinked Conzplex Between Acr3 H]phementioning

confidence: 99%

Photo‐Induced Crosslinking between Phenylalanine Transfer RNA and Messenger RNA on the Escherichia coli Ribosome

Matzke

Barta

Kuechler

1980

European Journal of Biochemistry

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Crosslinking between phenylalanyl transfer RNA from brewer's yeast and poly(U) can be obtained when ribosomal complexes are irradiated at > 300 nm; the ribosomal complexes were formed by enzymatic binding of the tRNA. The crosslinked product was isolated on oligo(dA)-cellulose columns. The experiments suggest that the photo-crosslinking occurs via the wybutine in the anticodon loop of the tRNA. This system can be used to study interactions between tRNA and messenger RNA on the ribosome.

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Section: Translocation Of the Crosslinked Conzplex Between Acr3 H]phementioning

confidence: 99%

Photo‐Induced Crosslinking between Phenylalanine Transfer RNA and Messenger RNA on the Escherichia coli Ribosome

Matzke

Barta

Kuechler

1980

European Journal of Biochemistry

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“…fMet-tRNA binding was assayed by the millipore filter technique [20] and was carried out in 50 mM Tris-HC1 pH 7.4, 7 mM MgAc,, 50 mM NH,CI and 1 mM 2-mercaptoethanol (buffer A). Formylmethionyl-puromycin formation was measured with the ethylacetate extraction procedure [21]. GTP hydrolysis was determined by extraction of the phosphomolybdate complex according to Kolakofsky et al [22].…”

Section: Initiution Assaysmentioning

confidence: 99%

The Involvement of Protein L 11 in the Joining of the 30‐S Initiation Complex to the 50‐S Subunit

Naaktgeboren¹,

Schrier²,

Möller³

et al. 1976

European Journal of Biochemistry

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Ribosomal protein L11 participates in the coupling of the 30-S initiation complex with the 5 0 3 subunit.P3, cores, lacking L7, L8, L12, L33, L10 and L11 were reconstituted with L7 and L10. These particles are unable to join successfully to the 30-S initiation complex, whereas reconstitution of the same cores in the presence of L7, L10 and L l l restores 60-80% of the original coupling activity. Po cores lacking only L7, L8, L12 and L33 are able to carry out one round of initiation, addition of L7 resulting in complete restoration of full activity. The data obtained with these P,, core particles resemble those obtained with untreated 50-S particles carrying thiostrepton, which prevents the binding of initiation factor IF-2 into the 70-S initiation complex.It is postulated that L11 induces a niche on the ribosomal surface to facilitate the proper binding of the IF-2 GTP 3 Met-tRNA complex. This binding of IF-2 enables the 30-S initiation complex to join to the 50-S subunit, because of the associative ability of IF-2. If joining is impaired then both the level of Met-tRNA binding and of the IF-Zmediated GTP hydrolysis is lowered.The physical association of both ribosomal subunits is a prerequisite for a number of reactions which are carried out by initiation and elongation factors and which are strictly ribosome-dependent. One of the most extensively studied processes is the hydrolysis of GTP mediated by the elongation factors EF-Tu and EF-G, in which it is known that several ribosomal proteins are involved (for a review see [l]). In the overall process of GTP hydrolysis L7 and L12 play an important role [2,3] by supplying the binding sites for these factors to the ribosomal surface [4,5] whereas proteins such as L5, L11 and L18 are more involved in the binding and the hydrolysis of GTP Far less has been discovered about the interaction of initiation factors with ribosomal proteins. It has already been established that L7 and L12 are needed for IF-2-mediated hydrolysis of GTP [lo, 111. Because, however, EF-Tu and IF-2 are mutually exclusive in 70-S initiation complexes, one can assume that initiation factors are also using the same or similar set of ribosomal proteins to fulfil their functions [12,13]. In this paper we have focussed our attention on possible roles of ribosomal proteins in the step in the 176, 195-197 (1972).

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“…In agreement with others (17) Leder and Bursztyn (23). Radioactivity was determined in 10 ml of a mixture containing Triton X-100/toluene/H20 (10:20:3) supplemented with 8.5 g of PPO (25).…”

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confidence: 53%

Specific in situ cleavage of 16S ribosomal RNA of Escherichia coli interferes with the function of initiation factor IF-1.

Baan¹,

Duijfjes²,

Leerdam³

et al. 1976

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

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Specific in situ cleavage of 16S rRNA of E.coli has been accomplished by in vitro treatment of 70S ribosomes ("tight couples") with the bacteriocin cloacin DF13. The defective ribosomes, which have fully lost their ability to sustain polypeptide synthesis, are still able to form initiation complexes with MS2 RNA, but the kinetics are altered. This is apparently due to an improper functioning of initiation factor IF-1, for the defective ribosomal couples respond normally to dissociation by IF-3 but the dissociation is not stimulated by IF-1. The initiation complexes formed with defective ribosomes are fully reactive with puromycin. Their ability to bind alanyl-tRNA is reduced by about 50% at all concentrations of elongation factor Tu studied. Cleavage of the 16S rRNA, not the release of the terminal fragment from the ribosome, causes the block of protein synthesis and the aberrations observed during initiation and elongation.The evidence is growing that the 3'-OH end of 16S rRNA of Escherichia coli is part of an important functional site on the 30S ribosome subunit. Specific cleavage of the 16S rRNA by the bacteriocin colicin ES, 49 nucleotides from the 3'-OH end, results in a total loss of the activity of the ribosome in translation (1). The presence of two adjacent methylated adenine residues near the 3' terminus of 16S rRNA determines sensitivity towards the antibiotic kasugamycin, a potent inhibitor of chain initiation (2). Crosslinking experiments indicate that a number of ribosomal proteins and initiation factors that have been implicated in initiation events, namely,

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Initiation of protein synthesis,I. Effect of formylation of methionyl-tRNA on codon recognition.

Cited by 34 publications

References 14 publications

Photo‐Induced Crosslinking between Phenylalanine Transfer RNA and Messenger RNA on the Escherichia coli Ribosome

Photo‐Induced Crosslinking between Phenylalanine Transfer RNA and Messenger RNA on the Escherichia coli Ribosome

The Involvement of Protein L 11 in the Joining of the 30‐S Initiation Complex to the 50‐S Subunit

Specific in situ cleavage of 16S ribosomal RNA of Escherichia coli interferes with the function of initiation factor IF-1.

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