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TitleStructure of the mammalian 80S initiation complex with initiation factor 5B on HCV-IRES RNA.
Journal, issue, pagesNat Struct Mol Biol, Vol. 21, Issue 8, Page 721-727, Year 2014
Publish dateJul 27, 2014
AuthorsHiroshi Yamamoto / Anett Unbehaun / Justus Loerke / Elmar Behrmann / Marianne Collier / Jörg Bürger / Thorsten Mielke / Christian M T Spahn /
PubMed AbstractThe universally conserved eukaryotic initiation factor (eIF) 5B, a translational GTPase, is essential for canonical translation initiation. It is also required for initiation facilitated by the ...The universally conserved eukaryotic initiation factor (eIF) 5B, a translational GTPase, is essential for canonical translation initiation. It is also required for initiation facilitated by the internal ribosomal entry site (IRES) of hepatitis C virus (HCV) RNA. eIF5B promotes joining of 60S ribosomal subunits to 40S ribosomal subunits bound by initiator tRNA (Met-tRNAi(Met)). However, the exact molecular mechanism by which eIF5B acts has not been established. Here we present cryo-EM reconstructions of the mammalian 80S-HCV-IRES-Met-tRNAi(Met)-eIF5B-GMPPNP complex. We obtained two substates distinguished by the rotational state of the ribosomal subunits and the configuration of initiator tRNA in the peptidyl (P) site. Accordingly, a combination of conformational changes in the 80S ribosome and in initiator tRNA facilitates binding of the Met-tRNAi(Met) to the 60S P site and redefines the role of eIF5B as a tRNA-reorientation factor.
External linksNat Struct Mol Biol / PubMed:25064512
MethodsEM (single particle)
Resolution8.9 - 9.5 Å
Structure data

2682

4ujd

EMDB-2682: Mammalian 80S-HCV-IRES initiation complex with eIF5B PRE-like state
PDB-4ujd: mammalian 80S HCV-IRES initiation complex with eIF5B PRE-like state
Method: EM (single particle) / Resolution: 8.9 Å

2683

4ujc

EMDB-2683: Mammalian 80S-HCV-IRES initiation complex with eIF5B POST-like state
PDB-4ujc: mammalian 80S HCV-IRES initiation complex with eIF5B POST-like state
Method: EM (single particle) / Resolution: 9.5 Å

Chemicals

ChemComp-GNP:
PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER / GppNHp, GMPPNP, energy-carrying molecule analogue*YM / 5'-Guanylyl imidodiphosphate

ChemComp-MG:
Unknown entry

Source
  • oryctolagus cuniculus (rabbit)
  • hepatitis c virus
KeywordsRIBOSOME / TRANSLATION INITIATION / HEPATITIS C VIRUS INTERNAL RIBOSOME ENTRY SITE / EUKARYOTIC INITIATION FACTOR 5B

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