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TitleStructural Insights into the Mammalian Late-Stage Initiation Complexes.
Journal, issue, pagesCell Rep, Vol. 31, Issue 1, Page 107497, Year 2020
Publish dateApr 7, 2020
AuthorsAngelita Simonetti / Ewelina Guca / Anthony Bochler / Lauriane Kuhn / Yaser Hashem /
PubMed AbstractIn higher eukaryotes, the mRNA sequence in the direct vicinity of the start codon, called the Kozak sequence (CRCCaugG, where R is a purine), is known to influence the rate of the initiation process. ...In higher eukaryotes, the mRNA sequence in the direct vicinity of the start codon, called the Kozak sequence (CRCCaugG, where R is a purine), is known to influence the rate of the initiation process. However, the molecular basis underlying its role remains poorly understood. Here, we present the cryoelectron microscopy (cryo-EM) structures of mammalian late-stage 48S initiation complexes (LS48S ICs) in the presence of two different native mRNA sequences, β-globin and histone 4, at overall resolution of 3 and 3.5 Å, respectively. Our high-resolution structures unravel key interactions from the mRNA to eukaryotic initiation factors (eIFs): 1A, 2, 3, 18S rRNA, and several 40S ribosomal proteins. In addition, we are able to study the structural role of ABCE1 in the formation of native 48S ICs. Our results reveal a comprehensive map of ribosome/eIF-mRNA and ribosome/eIF-tRNA interactions and suggest the impact of mRNA sequence on the structure of the LS48S IC.
External linksCell Rep / PubMed:32268096 / PubMed Central
MethodsEM (single particle)
Resolution3.0 - 14.3 Å
Structure data

10760

6yal

EMDB-10760, PDB-6yal:
Mammalian 48S late-stage initiation complex with beta-globin mRNA
Method: EM (single particle) / Resolution: 3.0 Å

10761

6yam

EMDB-10761, PDB-6yam:
Mammalian 48S late-stage translation initiation complex (LS48S+eIF3 IC) with beta-globin mRNA
Method: EM (single particle) / Resolution: 3.6 Å

10762

6yan

EMDB-10762, PDB-6yan:
Mammalian 48S late-stage translation initiation complex with histone 4 mRNA
Method: EM (single particle) / Resolution: 3.48 Å

EMDB-10763:
Mammalian late-stage translation initiation complex with beta-globin mRNA in presence of ATP
Method: EM (single particle) / Resolution: 9.6 Å

EMDB-10764:
Mammalian late-stage translation initiation complex with beta-globin mRNA in presence of ATP
Method: EM (single particle) / Resolution: 14.3 Å

Chemicals

ChemComp-SF4:
IRON/SULFUR CLUSTER / Iron–sulfur cluster

ChemComp-MG:
Unknown entry

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

Source
  • oryctolagus cuniculus (rabbit)
  • homo sapiens (human)
  • Rabbit (rabbit)
  • mus musculus (house mouse)
KeywordsTRANSLATION / initiation complex / 48S / eIF1A / eIF3 / ABCE1 / rabbit / translation initiation / eukaryotic initiation factor 1 / eukaryotic initiation factor 1A / eukaryotic initiation factor 3 / late-stage initiation complex

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