真核生物延伸因子3的一种新功能：抑制酵母中的终止密码子通读。
A novel function for eukaryotic elongation factor 3: Inhibition of stop codon readthrough in yeast.
作者信息
Kobayashi Soushi, Kaji Akira, Kaji Hideko
机构信息
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA, 19104, USA; Department of Biochemistry and Molecular Biology, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA, 19107, USA.
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA, 19104, USA.
出版信息
Arch Biochem Biophys. 2023 May 15;740:109580. doi: 10.1016/j.abb.2023.109580. Epub 2023 Mar 21.
Eukaryotic elongation factor 3 (eEF3) is one of the essential yeast ribosome-associated ATP-binding cassette type F (ABCF) ATPases. Previously, we found that eEF3 stimulates release of mRNA from puromycin-treated polysomes. In this study, we used a cell-free cricket paralysis virus (CrPV) internal ribosome entry site (IRES)-mediated firefly luciferase bicistronic mRNA translation system with yeast S30 extract. When eEF3 was partially removed from the crude extract, the product from the downstream ORF was increased by the readthrough of a UAA stop codon in the upstream ORF. eEF3 enhanced the release of luciferase from the polysome by eukaryotic release factor (eRF)1 and eRF3. These results suggest that eEF3 is a factor that assists eRFs in performing normal protein synthesis termination in yeast.
真核延伸因子3（eEF3）是酵母中一种必需的核糖体相关ATP结合盒F型（ABCF）ATP酶。此前，我们发现eEF3可刺激嘌呤霉素处理的多核糖体释放mRNA。在本研究中，我们使用了无细胞蟋蟀麻痹病毒（CrPV）内部核糖体进入位点（IRES）介导的萤火虫荧光素酶双顺反子mRNA翻译系统及酵母S30提取物。当从粗提物中部分去除eEF3时，上游开放阅读框（ORF）中的UAA终止密码子通读导致下游ORF的产物增加。eEF3通过真核释放因子（eRF）1和eRF3增强了荧光素酶从多核糖体的释放。这些结果表明，eEF3是一种辅助eRFs在酵母中进行正常蛋白质合成终止的因子。
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