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• W1788793928 startingPage "2896" @default.
• W1788793928 abstract "It has been shown previously [Brown, C.M. & Tate, W.P. (1994) J. Biol. Chem.269, 33164–33170.] that the polypeptide chain release factor RF2 involved in translation termination in prokaryotes was able to photocrossreact with mini-messenger RNAs containing stop signals in which U was replaced by 4-thiouridine (s4U). Here, using the same strategy we have monitored photocrosslinking to eukaryotic ribosomal components of 14-mer mRNA in the presence of , and 42-mer mRNA in the presence of tRNAAsp (tRNAAsp gene transcript). We show that: (a) both 14-mer and 42-mer mRNAs crossreact with ribosomal RNA and ribosomal proteins. The patterns of the crosslinked ribosomal proteins are similar with both mRNAs and sensitive to ionic conditions; (b) the crosslinking patterns obtained with 42-mer mRNAs show characteristic modification upon addition of tRNAAsp providing evidence for appropriate mRNA phasing onto the ribosome. Similar changes are not detected with the 14-mer pairs; (c) when eukaryotic polypeptide chain release factor 1 (eRF1) is added to the ribosome·tRNAAsp complex it crossreacts with the 42-mer mRNA containing the s4UGA stop codon located in the A site, but not with the s4UCA sense codon; this crosslink involves the N-terminal and middle domains of eRF1 but not the C domain which interacts with eukaryotic polypeptide chain release factor 3 (eRF3); (d) addition of eRF3 has no effect on the yield of eRF1–42-mer mRNA crosslinking and eRF3 does not crossreact with 42-mer mRNA. These experiments delineate the in vitro conditions allowing optimal phasing of mRNA on the eukaryotic ribosome and demonstrate a direct and specific contact of ‘core’ eRF1 and s4UGA stop codon within the ribosomal A site." @default.
• W1788793928 created "2016-06-24" @default.
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• W1788793928 date "2001-05-15" @default.
• W1788793928 modified "2023-09-30" @default.
• W1788793928 title "The polypeptide chain release factor eRF1 specifically contacts the s⁴UGA stop codon located in the A site of eukaryotic ribosomes" @default.
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• W1788793928 doi "https://doi.org/10.1046/j.1432-1327.2001.02177.x" @default.
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