Eukaryotic translation initiation factor 3 plays distinct roles at the mrna entry and exit channels of the ribosomal preinitiation complex

Colin Echeverrıá Aitken, Petra Beznosková, Vladislava Vlčkova, Wen Ling Chiu, Fujun Zhou, Leoš Shivaya Valásěk, Alan G. Hinnebusch, Jon R. Lorsch

Research output: Contribution to journalArticle

Abstract

Eukaryotic translation initiation factor 3 (eIF3) is a central player in recruitment of the pre-initiation complex (PIC) to mRNA. We probed the effects on mRNA recruitment of a library of S. cerevisiae eIF3 functional variants spanning its 5 essential subunits using an in vitro-reconstituted system. Mutations throughout eIF3 disrupt its interaction with the PIC and diminish its ability to accelerate recruitment to a native yeast mRNA. Alterations to the eIF3a CTD and eIF3b/i/g significantly slow mRNA recruitment, and mutations within eIF3b/i/g destabilize eIF2.GTP.Met-tRNAi binding to the PIC. Using model mRNAs lacking contacts with the 40S entry or exit channels, we uncovered a critical role for eIF3 requiring the eIF3a NTD, in stabilizing mRNA interactions at the exit channel, and an ancillary role at the entry channel requiring residues of the eIF3a CTD. These functions are redundant: defects at each channel can be rescued by filling the other channel with mRNA.

Original languageEnglish (US)
Article numbere20934
JournaleLife
Volume5
Issue numberOCTOBER2016
DOIs
StatePublished - Oct 26 2016
Externally publishedYes

Fingerprint

Prokaryotic Initiation Factor-3
Eukaryotic Initiation Factor-3
Eukaryotic Initiation Factors
Messenger RNA
Mutation
Guanosine Triphosphate
Yeast
Libraries
Saccharomyces cerevisiae
Yeasts
Defects

ASJC Scopus subject areas

  • Neuroscience(all)
  • Medicine(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Aitken, C. E., Beznosková, P., Vlčkova, V., Chiu, W. L., Zhou, F., Valásěk, L. S., ... Lorsch, J. R. (2016). Eukaryotic translation initiation factor 3 plays distinct roles at the mrna entry and exit channels of the ribosomal preinitiation complex. eLife, 5(OCTOBER2016), [e20934]. https://doi.org/10.7554/eLife.20934

Eukaryotic translation initiation factor 3 plays distinct roles at the mrna entry and exit channels of the ribosomal preinitiation complex. / Aitken, Colin Echeverrıá; Beznosková, Petra; Vlčkova, Vladislava; Chiu, Wen Ling; Zhou, Fujun; Valásěk, Leoš Shivaya; Hinnebusch, Alan G.; Lorsch, Jon R.

In: eLife, Vol. 5, No. OCTOBER2016, e20934, 26.10.2016.

Research output: Contribution to journalArticle

Aitken, CE, Beznosková, P, Vlčkova, V, Chiu, WL, Zhou, F, Valásěk, LS, Hinnebusch, AG & Lorsch, JR 2016, 'Eukaryotic translation initiation factor 3 plays distinct roles at the mrna entry and exit channels of the ribosomal preinitiation complex', eLife, vol. 5, no. OCTOBER2016, e20934. https://doi.org/10.7554/eLife.20934
Aitken, Colin Echeverrıá ; Beznosková, Petra ; Vlčkova, Vladislava ; Chiu, Wen Ling ; Zhou, Fujun ; Valásěk, Leoš Shivaya ; Hinnebusch, Alan G. ; Lorsch, Jon R. / Eukaryotic translation initiation factor 3 plays distinct roles at the mrna entry and exit channels of the ribosomal preinitiation complex. In: eLife. 2016 ; Vol. 5, No. OCTOBER2016.
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