gkz600.pdf (5.01 MB)
A role for the Saccharomyces cerevisiae ABCF protein New1 in translation termination/recycling
journal contribution
posted on 2023-06-09, 18:24 authored by Villu Kasari, Agnieszka A Pochopien, Tõnu Margus, Victoriia Murina, Kathryn Turnbull, Yang Zhou, Tracy Nissan, Michael Graf, Jirí Novácek, Gemma C Atkinson, Marcus J O Johansson, Daniel N Wilson, Vasili HauryliukTranslation is controlled by numerous accessory proteins and translation factors. In the yeast Saccharomyces cerevisiae, translation elongation requires an essential elongation factor, the ABCF ATPase eEF3. A closely related protein, New1, is encoded by a non-essential gene with cold sensitivity and ribosome assembly defect knock-out phenotypes. Since the exact molecular function of New1 is unknown, it is unclear if the ribosome assembly defect is direct, i.e. New1 is a bona fide assembly factor, or indirect, for instance due to a defect in protein synthesis. To investigate this, we employed yeast genetics, cryo-electron microscopy (cryo-EM) and ribosome profiling (Ribo-Seq) to interrogate the molecular function of New1. Overexpression of New1 rescues the inviability of a yeast strain lacking the otherwise strictly essential translation factor eEF3. The structure of the ATPase-deficient (EQ2) New1 mutant locked on the 80S ribosome reveals that New1 binds analogously to the ribosome as eEF3. Finally, Ribo-Seq analysis revealed that loss of New1 leads to ribosome queuing upstream of 3'-terminal lysine and arginine codons, including those genes encoding proteins of the cytoplasmic translational machinery. Our results suggest that New1 is a translation factor that fine-tunes the efficiency of translation termination or ribosome recycling.
History
Publication status
- Published
File Version
- Published version
Journal
Nucleic acids researchISSN
1362-4962Publisher
Oxford University PressExternal DOI
Issue
16Volume
47Page range
8807-8820Department affiliated with
- Biochemistry Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2019-07-15First Open Access (FOA) Date
2019-07-15First Compliant Deposit (FCD) Date
2019-07-15Usage metrics
Categories
No categories selectedKeywords
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC