The effects of codon context on in vivo translation speed.
The effects of codon context on in vivo translation speed.
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DOI：
10.1371/journal.pgen.1004392
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发表时间：
2014-06
期刊：
影响因子：
4.5
通讯作者：
Hughes KT
中科院分区：
文献类型：
--
作者：
Chevance FF;Le Guyon S;Hughes KT
我们开发了一个细菌遗传系统的基础上翻译的his操纵子前导肽基因，以确定相对速度的核糖体阅读单个或多个密码子在体内。所谓的“沉默”密码子变化的低频效应和密码子相邻（上下文）效应可以使用该测定法测量。该系统的优点是翻译速度不受His前导肽的一级序列的影响。我们表明，核糖体翻译同义密码子的表观速度可以有很大的不同，即使是相同的tRNA物种的同义密码子阅读。通过密码子对检测64个密码子相对于特定密码子的5′-和3′-侧定位的翻译，发现密码子对方向显著影响体内翻译速度。具有罕见精氨酸密码子和连续脯氨酸密码子的密码子对是体内翻译最慢的密码子对。该系统使我们能够确定不同因素对体内翻译速度的影响，包括Shine-Dalgarno序列，二肽键形成速率，密码子上下文和带电tRNA水平。分子生物学的中心法则指出，DNA转录成RNA，RNA翻译成蛋白质。通过核糖体将信使RNA（mRNA）翻译成蛋白质的过程是一个复杂的过程，涉及具有连接的氨基酸的转移RNA中间体，其必须识别mRNA序列中的3个碱基密码子以用正确的氨基酸翻译。用4个碱基编码20种蛋白质，遗传密码是冗余的，有61个编码密码子和3个终止密码子。一个给定的氨基酸可能由一个、两个、三个、四个或六个不同的密码子编码。我们开发了一个系统，可以测量核糖体在体内翻译过程中的速度。使用这个系统，我们表明，一个给定的密码子被翻译的能力是依赖于它的相邻密码子，密码子上下文效应。
We developed a bacterial genetic system based on translation of the his operon leader peptide gene to determine the relative speed at which the ribosome reads single or multiple codons in vivo. Low frequency effects of so-called “silent” codon changes and codon neighbor (context) effects could be measured using this assay. An advantage of this system is that translation speed is unaffected by the primary sequence of the His leader peptide. We show that the apparent speed at which ribosomes translate synonymous codons can vary substantially even for synonymous codons read by the same tRNA species. Assaying translation through codon pairs for the 5′- and 3′- side positioning of the 64 codons relative to a specific codon revealed that the codon-pair orientation significantly affected in vivo translation speed. Codon pairs with rare arginine codons and successive proline codons were among the slowest codon pairs translated in vivo. This system allowed us to determine the effects of different factors on in vivo translation speed including Shine-Dalgarno sequence, rate of dipeptide bond formation, codon context, and charged tRNA levels. The central dogma of molecular biology states that DNA is transcribed into RNA, which is translated into protein. The process of translation from messenger RNA (mRNA) into protein by the ribosome is a complicated process involving transfer RNA intermediates with attached amino acids that must recognize 3 base codons in the mRNA sequence to be translated with the correct amino acid. With 4 bases to code for 20 proteins the genetic code is redundant with 61 coding codons and three stop codons. A given amino acid might be coded by one, two, three, four or six different codons. We developed a system that measures the speed of the ribosome in vivo during translation. Using this system, we show that the ability of a given codon to be translated is dependent on its neighboring codons, the codon context effect.
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影响因子：
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作者：
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发表时间：
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