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New insights into stop codon recognition by eRF1

Blanchet, Sandra, Rowe, Michelle, von der Haar, Tobias, Fabret, Céline, Demais, Stéphane, Howard, Mark J., Namy, Olivier (2015) New insights into stop codon recognition by eRF1. Nucleic Acids Research, 43 (6). pp. 3298-3308. ISSN 0305-1048. (doi:10.1093/nar/gkv154) (KAR id:51589)

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http://dx.doi.org/10.1093/nar/gkv154

Abstract

In eukaryotes, translation termination is performed by eRF1, which recognizes stop codons via its N-terminal domain. Many previous studies based on point mutagenesis, cross-linking experiments or eRF1 chimeras have investigated the mechanism by which the stop signal is decoded by eRF1. Conserved motifs, such as GTS and YxCxxxF, were found to be important for termination efficiency, but the recognition mechanism remains unclear. We characterized a region of the eRF1 N-terminal domain, the P1 pocket, that we had previously shown to be involved in termination efficiency. We performed alanine scanning mutagenesis of this region, and we quantified in vivo readthrough efficiency for each alanine mutant. We identified two residues, arginine 65 and lysine 109, as critical for recognition of the three stop codons. We also demonstrated a role for the serine 33 and serine 70 residues in UGA decoding in vivo. NMR analysis of the alanine mutants revealed that the correct conformation of this region was controlled by the YxCxxxF motif. By combining our genetic data with a structural analysis of eRF1 mutants, we were able to formulate a new model in which the stop codon interacts with eRF1 through the P1 pocket.

Item Type: Article
DOI/Identification number: 10.1093/nar/gkv154
Subjects: Q Science > QP Physiology (Living systems) > QP506 Molecular biology
Q Science > QR Microbiology
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Tobias von der Haar
Date Deposited: 18 Jan 2017 12:52 UTC
Last Modified: 07 Feb 2020 15:53 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/51589 (The current URI for this page, for reference purposes)
von der Haar, Tobias: https://orcid.org/0000-0002-6031-9254
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