A role for the Saccharomyces cerevisiae ABCF protein New1 in translation termination/recycling
| Authors | |
|---|---|
| Year of publication | 2019 |
| Type | Article in Periodical |
| Magazine / Source | Nucleic acids research |
| MU Faculty or unit | |
| Citation | |
| Web | http://sro.sussex.ac.uk/id/eprint/84897/1/gkz600.pdf |
| Doi | http://dx.doi.org/10.1093/nar/gkz600 |
| Keywords | MESSENGER-RNA DECAY; INITIATION-FACTOR; GENE ONTOLOGY; YEAST; ELONGATION; COMPLEX; VISUALIZATION; PURIFICATION; BIOGENESIS; EXPRESSION |
| Description | 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 (EQ(2)) 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. |
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