Translational initiation factor eIF5 replaces eIF1 on the 40S ribosomal subunit to promote start-codon recognition.

Llácer, Jose Luis; Hussain, Tanweer; Saini, Adesh K; Nanda, Jagpreet Singh; Kaur, Sukhvir; Gordiyenko, Yuliya; Kumar, Rakesh; Hinnebusch, Alan G et al. · Elife · 2018

basic_science · Level V

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Abstract

In eukaryotic translation initiation, AUG recognition of the mRNA requires accommodation of Met-tRNA<sub>i</sub> in a 'P<sub>IN</sub>' state, which is antagonized by the factor eIF1. eIF5 is a GTPase activating protein (GAP) of eIF2 that additionally promotes stringent AUG selection, but the molecular basis of its dual function was unknown. We present a cryo-electron microscopy (cryo-EM) reconstruction of a yeast 48S pre-initiation complex (PIC), at an overall resolution of 3.0 Å, featuring the N-terminal domain (NTD) of eIF5 bound to the 40S subunit at the location vacated by eIF1. eIF5 interacts with and allows a more accommodated orientation of Met-tRNA<sub>i</sub>. Substitutions of eIF5 residues involved in the eIF5-NTD/tRNA<sub>i</sub> interaction influenced initiation at near-cognate UUG codons<i>in vivo,</i> and the closed/open PIC conformation in vitro, consistent with direct stabilization of the codon:anticodon duplex by the wild-type eIF5-NTD. The present structure reveals the basis for a key role of eIF5 in start-codon selection.

Medical subject headings