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一种与非 AUG 密码子翻译起始信号有关的病毒 RNA 基序。

A viral RNA motif involved in signaling the initiation of translation on non-AUG codons.

机构信息

Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco 28049 Madrid, Spain.

出版信息

RNA. 2019 Apr;25(4):431-452. doi: 10.1261/rna.068858.118. Epub 2019 Jan 18.

DOI:10.1261/rna.068858.118
PMID:30659060
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6426287/
Abstract

Noncanonical translation, and particularly initiation on non-AUG codons, are frequently used by viral and cellular mRNAs during virus infection and disease. The Sindbis virus (SINV) subgenomic mRNA (sgRNA) constitutes a unique model system to analyze the translation of a capped viral mRNA without the participation of several initiation factors. Moreover, sgRNA can initiate translation even when the AUG initiation codon is replaced by other codons. Using SINV replicons, we examined the efficacy of different codons in place of AUG to direct the synthesis of the SINV capsid protein. The substitution of AUG by CUG was particularly efficient in promoting the incorporation of leucine or methionine in similar percentages at the amino terminus of the capsid protein. Additionally, valine could initiate translation when the AUG is replaced by GUG. The ability of sgRNA to initiate translation on non-AUG codons was dependent on the integrity of a downstream stable hairpin (DSH) structure located in the coding region. The structural requirements of this hairpin to signal the initiation site on the sgRNA were examined in detail. Of interest, a virus bearing CUG in place of AUG in the sgRNA was able to infect cells and synthesize significant amounts of capsid protein. This virus infects the human haploid cell line HAP1 and the double knockout variant that lacks eIF2A and eIF2D. Collectively, these findings indicate that leucine-tRNA or valine-tRNA can participate in the initiation of translation of sgRNA by a mechanism dependent on the DSH. This mechanism does not involve the action of eIF2, eIF2A, or eIF2D.

摘要

非规范翻译，特别是起始于非 AUG 密码子，在病毒感染和疾病期间，经常被病毒和细胞 mRNA 所使用。辛德毕斯病毒（SINV）亚基因组 mRNA（sgRNA）构成了一个独特的模型系统，可用于分析无几个起始因子参与的有帽病毒 mRNA 的翻译。此外，即使 AUG 起始密码子被其他密码子取代，sgRNA 也可以起始翻译。使用 SINV 复制子，我们研究了不同密码子取代 AUG 以指导 SINV 衣壳蛋白合成的效率。用 CUG 取代 AUG 特别有效地促进了亮氨酸或甲硫氨酸以相似的百分比掺入衣壳蛋白的氨基末端。此外，当 AUG 被 GUG 取代时，缬氨酸也可以起始翻译。sgRNA 能够在非 AUG 密码子上起始翻译取决于位于编码区的下游稳定发夹（DSH）结构的完整性。详细研究了该发夹结构启动 sgRNA 起始位点的结构要求。有趣的是，携带 sgRNA 中 AUG 被 CUG 取代的病毒能够感染细胞并合成大量衣壳蛋白。这种病毒感染人类单倍体细胞系 HAP1 和缺乏 eIF2A 和 eIF2D 的双敲除变体。总之，这些发现表明亮氨酰-tRNA 或缬氨酰-tRNA 可以通过依赖于 DSH 的机制参与 sgRNA 翻译的起始。该机制不涉及 eIF2、eIF2A 或 eIF2D 的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/e1873760ce77/431f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/ed243070d1dd/431f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/2b0fbd3afbf6/431f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/f7c4d472cced/431f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/7fec5152db5a/431f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/ff8dea47c007/431f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/7c6762ab01d1/431f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/b01891941e91/431f07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/977d89ea8c13/431f08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/562f6e25e892/431f09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/e1873760ce77/431f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/ed243070d1dd/431f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/2b0fbd3afbf6/431f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/f7c4d472cced/431f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/7fec5152db5a/431f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/ff8dea47c007/431f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/7c6762ab01d1/431f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/b01891941e91/431f07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/977d89ea8c13/431f08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/562f6e25e892/431f09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8f7/6426287/e1873760ce77/431f10.jpg

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