The rescue of stalled translational conplexes: recoding of a sense to nonsense codon

拯救停滞的翻译复合体：将有义密码子重新编码为无义密码子

• 批准号: BB/E010709/1
• 负责人: Martin Ryan
• 金额: $ 45.78万
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE010709%2F1
• 关键词: rescue; stalled; translational; conplexes; recoding

Ribosomes synthesise all the proteins in the cell by reading or 'translating' information in messenger RNA (mRNA), which in turn is a copy of information stored in the cell's DNA. Accurate translation by ribosomes relies on them obeying certain rules, the genetic code, in which each triplet of nucleotides is always read in the same way to initiate, continue or terminate synthesis of a completed protein. The fidelity of translation is extremely high, and very few errors are made. Surprisingly however, on occasion ribosomes are prompted to disregard this code, often by particular sequences in the mRNA that is being read or the protein sequence that has just been synthesized by the ribosome and is still inside it. Such 'recoding' events are often found in viral RNAs and allow viruses to generate all the proteins they need from very compact, efficient genomes. Some recoding events also take place on cellular mRNAs and can be very important for correct expression of proteins. In many cases recoding events allow the normal 'stop' signals that signify the end of a protein to be bypassed, leading to extension of the protein. We have uncovered a novel recoding event in which the ribosome is prompted to stop translation and then restart / without the normal signals for either / thereby generating 2 separate proteins from one mRNA. This is dictated by a short peptide sequences termed '2A' from viruses and provides both an important and convenient tool for co-expression of more than one protein without the need for multiple mRNAs, and also the possibility of insight into how the ribosome works. Understanding this event may have another important repercussions as it may allow development of antiviral strategies, aimed at inhibiting this recoding event during viral infection. Our aim is therefore to understand reaction dictated by 2A in detail, identify all the factors that are required and the cellular functions that it impinges on. Thus far we have found that the 2A peptide causes ribosomes to pause, and that the 'release factors' that normally catalyse termination of translation at a stop signal are required for the abnormal termination reaction that takes place at 2A. We will investigate the interactions of release factors with ribosomes paused at 2A and attempt to determine what factors contribute to the pause.

核糖体通过阅读或“翻译”信使RNA（mRNA）中的信息来合成细胞中的所有蛋白质，而信使RNA又是储存在细胞DNA中的信息的副本。核糖体的准确翻译依赖于它们遵守某些规则，即遗传密码，其中每个核苷酸三联体总是以相同的方式阅读，以启动，继续或终止完整蛋白质的合成。翻译的忠实度极高，很少出错。然而，令人惊讶的是，有时核糖体会忽略这一密码，通常是由正在阅读的mRNA中的特定序列或核糖体刚刚合成并仍在其内部的蛋白质序列引起的。这种“重新编码”事件通常在病毒RNA中发现，并允许病毒从非常紧凑，高效的基因组中产生它们需要的所有蛋白质。一些重编码事件也发生在细胞mRNA上，并且对于蛋白质的正确表达非常重要。在许多情况下，重新编码事件允许绕过表示蛋白质末端的正常“停止”信号，从而导致蛋白质的延伸。我们已经发现了一种新的重新编码事件，其中核糖体被提示停止翻译，然后重新启动/没有正常的信号，从而从一个mRNA产生2个独立的蛋白质。这是由来自病毒的称为“2A”的短肽序列决定的，并且提供了用于共表达多于一种蛋白质而不需要多个mRNA的重要且方便的工具，并且还提供了洞察核糖体如何工作的可能性。了解这一事件可能有另一个重要的影响，因为它可能允许开发抗病毒策略，旨在抑制病毒感染期间的这种重新编码事件。因此，我们的目标是详细了解由2A决定的反应，确定所需的所有因子及其影响的细胞功能。到目前为止，我们已经发现2A肽导致核糖体暂停，并且在2A处发生的异常终止反应需要通常催化终止翻译的“释放因子”。我们将研究释放因子与核糖体在2A处暂停的相互作用，并试图确定哪些因素有助于暂停。

项目成果

Characterization of native protein complexes and protein isoform variation using size-fractionation-based quantitative proteomics.

• DOI: 10.1074/mcp.m113.032367
• 发表时间: 2013-12
• 期刊: Molecular & cellular proteomics : MCP
• 作者: Kirkwood KJ;Ahmad Y;Larance M;Lamond AI
• 通讯作者: Lamond AI

Both cis and trans Activities of Foot-and-Mouth Disease Virus 3D Polymerase Are Essential for Viral RNA Replication.

• DOI: 10.1128/jvi.00469-16
• 发表时间: 2016-08-01
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Herod MR;Ferrer-Orta C;Loundras EA;Ward JC;Verdaguer N;Rowlands DJ;Stonehouse NJ
• 通讯作者: Stonehouse NJ

Genetic economy in picornaviruses: Foot-and-mouth disease virus replication exploits alternative precursor cleavage pathways.

• DOI: 10.1371/journal.ppat.1006666
• 发表时间: 2017-10
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Herod MR;Gold S;Lasecka-Dykes L;Wright C;Ward JC;McLean TC;Forrest S;Jackson T;Tuthill TJ;Rowlands DJ;Stonehouse NJ
• 通讯作者: Stonehouse NJ

Evaluating the use of HILIC in large-scale, multi dimensional proteomics: Horses for courses?

• DOI: 10.1016/j.ijms.2015.07.029
• 发表时间: 2015-11-30
• 期刊: International journal of mass spectrometry
• 影响因子: 1.8
• 作者: Bensaddek D;Nicolas A;Lamond AI
• 通讯作者: Lamond AI

A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells.

• DOI: 10.7554/elife.01630
• 发表时间: 2014-01-01
• 期刊: eLife
• 影响因子: 7.7
• 作者: Ly T;Ahmad Y;Shlien A;Soroka D;Mills A;Emanuele MJ;Stratton MR;Lamond AI
• 通讯作者: Lamond AI