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Protein S3 fragments neighboring mRNA during elongation and termination of translation on the human ribosome

Item Type:Article
Title:Protein S3 fragments neighboring mRNA during elongation and termination of translation on the human ribosome
Creators Name:Khairulina, Y.S. and Molotkov, M.V. and Bulygin, K.N. and Graifer, D.M. and Ven’yaminova, A.G. and Frolova, L.Y. and Stahl, J. and Karpova, G.G.
Abstract:Protein S3 fragments were determined that crosslink to modified mRNA analogues in positions +5 to +12 relative to the first nucleotide in the P-site bound codon in model complexes mimicking states of ribosomes at the elongation and translation termination steps. The mRNA analogues contained a Phe codon UUU/UUC at the 5′-termini that could predetermine the position of the tRNAPhe on the ribosome by the P-site binding and perfluorophenylazidobenzoyl group at a nucleotide in various positions 3′ of the UUU/UUC codon. The crosslinked S3 protein was isolated from 80S ribosomal complexes irradiated with mild UV light and subjected to cyanogen bromide—induced cleavage at methionine residues with subsequent identification of the crosslinked oligopeptides. An analysis of the positions of modified oligopeptides resulting from the cleavage showed that, in dependence on the positions of modified nucleotides in the mRNA analogue, the crosslinking sites were found in the N-terminal half of the protein (fragment 2–217) and/or in the C-terminal fragment 190–236; the latter reflects a new peculiarity in the structure of the mRNA binding center in the ribosome, unknown to date. The results of crosslinking did not depend on the type of A-site codon or on the presence of translation termination factor eRF1.
Keywords:Ribosome, Photoaffinity Cross-Linking, Eukaryotes, mRNA Analogue, mRNA Binding Site, Ribosomal Protein S3
Source:Russian Journal of Bioorganic Chemistry
ISSN:1068-1620
Publisher:Springer (Germany)
Volume:34
Number:6
Page Range:691-697
Date:November 2008
Official Publication:https://doi.org/10.1134/S1068162008060071
PubMed:View item in PubMed

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