New insights into stop codon recognition by eRF1

Nucleic Acids Research
Sandra BlanchetOlivier Namy

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.

References

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Citations

Sep 19, 2015·Nucleic Acids Research·Sarah MatheislRoland Beckmann
Nov 17, 2015·Life·Ana R BezerraManuel A S Santos
Dec 24, 2016·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Elena Alkalaeva, Tatiana Mikhailova
May 8, 2018·Cold Spring Harbor Perspectives in Biology·Christopher U T Hellen
May 18, 2016·Genetics·Thomas E DeverGraham D Pavitt
Jan 5, 2018·Proceedings of the National Academy of Sciences of the United States of America·Thomas Philipp HoernesMatthias David Erlacher
Oct 24, 2019·Human Genetics·Dora Janeth FonsecaPaul Laissue
Feb 12, 2020·International Journal of Molecular Sciences·Christian BeißelHeike Krebber
Jul 10, 2020·Journal of Neurochemistry·Debra AbramovJacqueline Burré
Feb 15, 2017·BMC Biology·Tomáš PánekMarek Eliáš
Mar 23, 2021·Cellular and Molecular Life Sciences : CMLS·Patrícia Martins-Dias, Luísa Romão
Jul 24, 2020·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Rachida AmzalEmmanuel Gonzales

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Methods Mentioned

BETA
X-ray
nuclear magnetic resonance
NMR
electrophoresis

Software Mentioned

NMRPipe
CCPN Analysis
Bruker TopSpin
R

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