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

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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

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