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PreprintCited by 80RNA and protein synthesis mechanismsRNA modifications and cancerPI3K/AKT/mTOR signaling in cancer

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eIF5B and eIF1A remodel human translation initiation complexes to mediate ribosomal subunit joining

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Lapointe, Christopher P., Grosely, Rosslyn, Sokabe, Masaaki, Alvarado, Carlos, Wang, Jinfan, Montabana, Elizabeth, Villa, Nancy, Shin, Byung-Sik, Dever, Thomas E., Fraser, Christopher S., Fernández, Israel S., Puglisi, Joseph D.·2021·Nature
DOI 10.1101/2021.12.09.471821PMC PMC9728550

Preprint — published as 10.1038/s41586-022-04858-z · Paper page

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Abstract

Joining of the ribosomal subunits at a translation start site on a messenger RNA during initiation commits the ribosome to synthesize a protein. Here, we combined single-molecule spectroscopy and structural methods using an in vitro reconstituted system to examine how the human ribosomal subunits join. Single-molecule fluorescence revealed when universally-conserved eukaryotic initiation factors (eIFs) eIF1A and eIF5B associate with and depart from initiation complexes. Guided by single-molecule dynamics, we examined initiation complexes that contained both eIF1A and eIF5B using single-particle electron cryo-microscopy. The resulting structure illuminated how eukaryote-specific contacts between eIF1A and eIF5B remodel the initiation complex to orient initiator tRNA in a conformation compatible with ribosomal subunit joining. Collectively, our findings provide a quantitative and architectural framework for the molecular choreography orchestrated by eIF1A and eIF5B during human translation initiation.

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Metadata

Authors
Lapointe, C. P., Grosely, R., Sokabe, M., Alvarado, C., Wang, J., Montabana, E., Villa, N., Shin, B.-S., Dever, T. E., Fraser, C. S., Fernández, I. S., & Puglisi, J. D.
Affiliations
  • Stanford Medicine (US)
  • University of California, Davis (US)
  • National Institutes of Health (US)
  • St. Jude Children's Research Hospital (US)
Corresponding author
  • Fernández, Israel S.
  • Puglisi, Joseph D.
ORCID
Venue
Nature
Publisher
openRxiv
Published
2021-12-09
Volume
607
Issue
7917
Pages
185-190
ISSN
0028-0836 (ISSN-L)
DOI
10.1101/2021.12.09.471821
PMC
PMC9728550
OpenAlex
W4283259136
Cited by
80
References
1
Citations by year
2026: 16 · 2025: 21 · 2024: 21 · 2023: 16 · 2022: 6
FWCI
5.22
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Nature (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • Damon Runyon Cancer Research Foundation (2321-18, DRG#2321-18)
  • Knut och Alice Wallenbergs Stiftelse (KAW 2015.0406, 2015.0406)
  • National Institutes of Health (GM011378, AG064690, AI047365)
  • Stanford Bio-X
  • National Institute on Aging (RF1AG064690, R01AG064690)
  • National Institute of General Medical Sciences (R01GM113078, R00GM144678, R01GM051266, K99GM144678, R01GM092927, R35GM145306)
  • SLAC National Accelerator Laboratory
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
Keywords
translation initiation, eIF1A, eIF5B, initiator tRNA, initiation complex, eukaryotic initiation factors, ribosome assembly, single-molecule fluorescence, single-particle cryo-electron microscopy
MeSH terms
Single Molecule Imaging, Humans, RNA, Transfer, Met, Eukaryotic Initiation Factor-1, Cryoelectron Microscopy, Eukaryotic Initiation Factors, Ribosome Subunits

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