A role for the Saccharomyces cerevisiae ABCF protein New1 in translation termination/recycling

Kasari, Villu, Pochopien, Agnieszka A, Margus, Tõnu, Murina, Victoriia, Turnbull, Kathryn, Zhou, Yang, Nissan, Tracy, Graf, Michael, Nováček, Jiří, Atkinson, Gemma C, Johansson, Marcus J O, Wilson, Daniel N and Hauryliuk, Vasili (2019) A role for the Saccharomyces cerevisiae ABCF protein New1 in translation termination/recycling. Nucleic acids research, 47 (16). pp. 8807-8820. ISSN 1362-4962

[img] PDF - Published Version
Available under License Creative Commons Attribution.

Download (5MB)

Abstract

Translation 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.

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
Subjects: Q Science > QR Microbiology > QR0001 General
Depositing User: Tracy Nissan
Date Deposited: 15 Jul 2019 08:21
Last Modified: 02 Oct 2020 14:30
URI: http://sro.sussex.ac.uk/id/eprint/84897

View download statistics for this item

📧 Request an update