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Structure paper

TitleTranslational initiation factor eIF5 replaces eIF1 on the 40S ribosomal subunit to promote start-codon recognition.
Journal, issue, pagesElife, Vol. 7, Year 2018
Publish dateNov 30, 2018
AuthorsJose Luis Llácer / Tanweer Hussain / Adesh K Saini / Jagpreet Singh Nanda / Sukhvir Kaur / Yuliya Gordiyenko / Rakesh Kumar / Alan G Hinnebusch / Jon R Lorsch / V Ramakrishnan /
PubMed AbstractIn eukaryotic translation initiation, AUG recognition of the mRNA requires accommodation of Met-tRNA in a 'P' state, which is antagonized by the factor eIF1. eIF5 is a GTPase activating protein (GAP) ...In eukaryotic translation initiation, AUG recognition of the mRNA requires accommodation of Met-tRNA in a 'P' 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. Substitutions of eIF5 residues involved in the eIF5-NTD/tRNA interaction influenced initiation at near-cognate UUG codons 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.
External linksElife / PubMed:30475211 / PubMed Central
MethodsEM (single particle)
Resolution3.02 - 3.53 Å
Structure data

4327

6fyx

EMDB-4327: Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (Map C1)
PDB-6fyx: Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (model C1)
Method: EM (single particle) / Resolution: 3.5 Å

4328

6fyy

EMDB-4328: Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (Map 1)
PDB-6fyy: Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (model C2)
Method: EM (single particle) / Resolution: 3.02 Å

EMDB-4329:
Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (Map C2)
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-4330:
Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (Map A)
Method: EM (single particle) / Resolution: 3.53 Å

EMDB-4331:
Structure of a partial yeast 48S preinitiation complex with eIF5 N-terminal domain (Map B)
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-MET:
METHIONINE / Methionine

ChemComp-GCP:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-PCP, energy-carrying molecule analogue*YM

Source
  • kluyveromyces lactis (strain atcc 8585 / cbs 2359 / dsm 70799 / nbrc 1267 / nrrl y-1140 / wm37) (yeast)
  • saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • Kluyveromyces lactis (yeast)
KeywordsRIBOSOME / translation / initiation factors / 40S / eIF1A / eIF3 / eIF2 / eIF5 / tRNAi / 48S PIC / small ribosome subunit

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