Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation

Chaomin Sun, Jordi Querol-Audí, Stefanie A. Mortimer, Ernesto Arias-Palomo, Jennifer A. Doudna, Eva Nogales, Jamie H D Cate

Research output: Contribution to journalArticle

Abstract

The initiation of protein synthesis plays an essential regulatory role in human biology. At the center of the initiation pathway, the 13-subunit eukaryotic translation initiation factor 3 (eIF3) controls access of other initiation factors and mRNA to the ribosome by unknown mechanisms. Using electron microscopy (EM), bioinformatics and biochemical experiments, we identify two highly conserved RNA-binding motifs in eIF3 that direct translation initiation from the hepatitis C virus internal ribosome entry site (HCV IRES) RNA. Mutations in the RNA-binding motif of subunit eIF3a weaken eIF3 binding to the HCV IRES and the 40S ribosomal subunit, thereby suppressing eIF2-dependent recognition of the start codon. Mutations in the eIF3c RNA-binding motif also reduce 40S ribosomal subunit binding to eIF3, and inhibit eIF5B-dependent steps downstream of start codon recognition. These results provide the first connection between the structure of the central translation initiation factor eIF3 and recognition of the HCV genomic RNA start codon, molecular interactions that likely extend to the human transcriptome.

Original languageEnglish (US)
Pages (from-to)7512-7521
Number of pages10
JournalNucleic Acids Research
Volume41
Issue number15
DOIs
StatePublished - Aug 2013
Externally publishedYes

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Prokaryotic Initiation Factor-3
Eukaryotic Initiation Factor-3
Eukaryotic Initiation Factors
Hepacivirus
Initiator Codon
Eukaryotic Small Ribosome Subunits
Peptide Initiation Factors
RNA
Mutation
Computational Biology
Ribosomes
Transcriptome
Electron Microscopy
RNA-Binding Motifs
Internal Ribosome Entry Sites
Messenger RNA

ASJC Scopus subject areas

  • Genetics

Cite this

Sun, C., Querol-Audí, J., Mortimer, S. A., Arias-Palomo, E., Doudna, J. A., Nogales, E., & Cate, J. H. D. (2013). Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation. Nucleic Acids Research, 41(15), 7512-7521. https://doi.org/10.1093/nar/gkt510

Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation. / Sun, Chaomin; Querol-Audí, Jordi; Mortimer, Stefanie A.; Arias-Palomo, Ernesto; Doudna, Jennifer A.; Nogales, Eva; Cate, Jamie H D.

In: Nucleic Acids Research, Vol. 41, No. 15, 08.2013, p. 7512-7521.

Research output: Contribution to journalArticle

Sun, C, Querol-Audí, J, Mortimer, SA, Arias-Palomo, E, Doudna, JA, Nogales, E & Cate, JHD 2013, 'Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation', Nucleic Acids Research, vol. 41, no. 15, pp. 7512-7521. https://doi.org/10.1093/nar/gkt510
Sun C, Querol-Audí J, Mortimer SA, Arias-Palomo E, Doudna JA, Nogales E et al. Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation. Nucleic Acids Research. 2013 Aug;41(15):7512-7521. https://doi.org/10.1093/nar/gkt510
Sun, Chaomin ; Querol-Audí, Jordi ; Mortimer, Stefanie A. ; Arias-Palomo, Ernesto ; Doudna, Jennifer A. ; Nogales, Eva ; Cate, Jamie H D. / Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation. In: Nucleic Acids Research. 2013 ; Vol. 41, No. 15. pp. 7512-7521.
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