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TitleThe assembly factor Reh1 is released from the ribosome during its initial round of translation.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 1278, Year 2025
Publish dateFeb 3, 2025
AuthorsSharmishtha Musalgaonkar / James N Yelland / Ruta Chitale / Shilpa Rao / Hakan Ozadam / David W Taylor / Can Cenik / Arlen W Johnson /
PubMed AbstractAssembly of functional ribosomal subunits and successfully delivering them to the translating pool is a prerequisite for protein synthesis and cell growth. In S. cerevisiae, the ribosome assembly ...Assembly of functional ribosomal subunits and successfully delivering them to the translating pool is a prerequisite for protein synthesis and cell growth. In S. cerevisiae, the ribosome assembly factor Reh1 binds to pre-60S subunits at a late stage during their cytoplasmic maturation. Previous work shows that the C-terminus of Reh1 inserts into the polypeptide exit tunnel of the pre-60S subunit. Here, we show that Reh1-bound nascent 60S subunits associate with 40S subunits to form actively translating ribosomes. Using selective ribosome profiling, we found that Reh1-bound ribosomes populate open reading frames near start codons. Reh1-bound ribosomes are also strongly enriched for initiator tRNA, indicating they are associated with early elongation. Using cryo-electron microscopy to image Reh1-bound 80S ribosomes, we found they contain A site peptidyl tRNA, P site tRNA and eIF5A, indicating that Reh1 does not dissociate from 60S until translation elongation. We propose that Reh1 is displaced by the elongating peptide chain, making it the last assembly factor released from the nascent 60S subunit during its initial round of translation.
External linksNat Commun / PubMed:39900920 / PubMed Central
MethodsEM (single particle)
Resolution3.13 - 3.22 Å
Structure data

42525

8ut0

EMDB-42525, PDB-8ut0:
Eukaryotic 80S ribosome with Reh1, eIF5A and A/P site tRNA
Method: EM (single particle) / Resolution: 3.22 Å

42540

8uti

EMDB-42540, PDB-8uti:
Eukaryotic 80S ribosome with Reh1 and A/P site tRNA
Method: EM (single particle) / Resolution: 3.13 Å

Chemicals

ChemComp-3HE:
4-{(2R)-2-[(1S,3S,5S)-3,5-dimethyl-2-oxocyclohexyl]-2-hydroxyethyl}piperidine-2,6-dione

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsRIBOSOME / ribosome assembly / translation

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