New insights into the incorporation of natural suppressor tRNAs at stop codons in Saccharomyces cerevisiae.
New insights into the incorporation of natural suppressor tRNAs at stop codons in Saccharomyces cerevisiae.
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DOI：
10.1093/nar/gku663
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发表时间：
2014-09
影响因子：
14.9
通讯作者：
Namy O
中科院分区：
文献类型：
--
作者：
Blanchet S;Cornu D;Argentini M;Namy O
核苷酸环境或药物可促进终止密码子的通读。在这种情况下，核糖体在终止密码子上结合了一个天然的抑制子tRNA，导致在同一阅读框架中继续翻译，直到下一个终止密码子，并导致具有新的潜在功能的蛋白质的表达。然而，参与终止密码子通读的天然抑制物tRNAs的身份仍然不清楚，这排除了识别参与终止位置的氨基酸的可能性。我们建立了一个体内报告系统，用于通过质谱仪鉴定酵母中包含在终止密码子上的氨基酸。我们发现谷氨酰胺、酪氨酸和赖氨酸在UAA和UAG密码子上插入，色氨酸、半胱氨酸和精氨酸在UGA密码子上插入。终止密码子的5‘核苷酸环境对通读结合的氨基酸的同一性或比例没有影响。我们还发现，两种不同的谷氨酰胺tRNAGln被用来在UAA和UAG密码子上插入谷氨酰胺。这项工作构成了对终止密码子结合的氨基酸的第一次系统分析，为核糖体用于读取遗传密码的解码规则提供了重要的新见解。
Stop codon readthrough may be promoted by the nucleotide environment or drugs. In such cases, ribosomes incorporate a natural suppressor tRNA at the stop codon, leading to the continuation of translation in the same reading frame until the next stop codon and resulting in the expression of a protein with a new potential function. However, the identity of the natural suppressor tRNAs involved in stop codon readthrough remains unclear, precluding identification of the amino acids incorporated at the stop position. We established an in vivo reporter system for identifying the amino acids incorporated at the stop codon, by mass spectrometry in the yeast Saccharomyces cerevisiae. We found that glutamine, tyrosine and lysine were inserted at UAA and UAG codons, whereas tryptophan, cysteine and arginine were inserted at UGA codon. The 5′ nucleotide context of the stop codon had no impact on the identity or proportion of amino acids incorporated by readthrough. We also found that two different glutamine tRNAGln were used to insert glutamine at UAA and UAG codons. This work constitutes the first systematic analysis of the amino acids incorporated at stop codons, providing important new insights into the decoding rules used by the ribosome to read the genetic code.
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核糖体分析揭示了果蝇果蝇中普遍存在的终止密码子读取。
影响因子：
7.7
作者：
Dunn JG;Foo CK;Belletier NG;Gavis ER;Weissman JS
通讯作者：
Weissman JS
影响因子：
14.9
作者：
Namy O;Duchateau-Nguyen G;Hatin I;Hermann-Le Denmat S;Termier M;Rousset JP
通讯作者：
Rousset JP
影响因子：
3.2
作者：
BOUADLOUN, F;SRICHAIYO, T;BJORK, GR
通讯作者：
BJORK, GR
影响因子：
5.6
作者：
HIRSH
通讯作者：
HIRSH
影响因子：
4.5
作者：
Bertram, G;Bell, HA;Stansfield, I
通讯作者：
Stansfield, I