A comprehensive analysis of yeast Elongator phosphorylation and its functional consequences

酵母Elongator磷酸化及其功能后果的综合分析

• 批准号: BB/F019629/1
• 负责人: Michael Stark
• 金额: $ 43.27万
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF019629%2F1
• 关键词: comprehensive; analysis; yeast; Elongator; phosphorylation

The addition and removal of phosphate groups at specific points within protein molecules ('protein phosphorylation') is a universal means by which protein function can be regulated. Phosphate groups are added by enzymes termed protein kinases and removed by opposing activities termed protein phosphatases. We are interested in a complex of proteins termed 'Elongator' that is found in all higher cells including those of plants and animals such as ourselves. We have been using yeast cells as a model system for these studies because they provide an experimental system in which both genetic and biochemical approaches to biological questions can be combined in a particularly powerful way. Elongator has been proposed to play a number of cellular roles but most recently it has been discovered that it is important for the correct functioning of transfer RNAs, special molecules that are required for cells to make more protein molecules using their genetic templates, and this is probably Elongator's primary function. We have found that one of the components of the Elongator complex appears to be regulated by phosphorylation, even though this might mean that Elongator is not fully active under all circumstances. Thus the cell's ability to synthesis specific proteins may be controlled through this mechanism. In our proposed work we will combine the expertise of our two laboratories to study both how and why Elongator may be regulated by phosphorylation. We will examine whether changes in Elongator activity lead to effects on the synthesis of specific proteins in the cell and identify both the protein kinases involved in phosphorylating Elongator and the specific sites in the component proteins of Elongator that are modified by these protein kinases. By defining individual sites of phosphorylation we can generate mutants that either cannot be phosphorylated at all or that mimic the presence of a phosphate group. We will use such mutants to build up a detailed picture of how specific phosphhorylation events affect Elongator function.

在蛋白质分子中特定点上磷酸基团的添加和去除（“蛋白质磷酸化”）是调节蛋白质功能的通用手段。磷酸基团由称为蛋白激酶的酶添加，并通过称为蛋白磷酸酶的相反活性去除。我们感兴趣的是一种名为“伸长体”的蛋白质复合物，它存在于所有高等细胞中，包括植物和动物的细胞，比如我们自己。我们一直使用酵母细胞作为这些研究的模型系统，因为它们提供了一个实验系统，在这个实验系统中，遗传和生化方法可以以一种特别强大的方式结合在一起。细长子被认为在细胞中发挥着许多作用，但最近发现它对转移rna的正确功能很重要，转移rna是细胞使用其遗传模板制造更多蛋白质分子所需的特殊分子，这可能是细长子的主要功能。我们发现，拉长复合体的一个组成部分似乎受到磷酸化的调节，尽管这可能意味着拉长复合体在所有情况下都不完全活跃。因此，细胞合成特定蛋白质的能力可能通过这种机制来控制。在我们提出的工作中，我们将结合我们两个实验室的专业知识来研究伸长子如何以及为什么可能受到磷酸化的调节。我们将研究伸长体活性的变化是否会对细胞中特定蛋白质的合成产生影响，并确定参与磷酸化伸长体的蛋白激酶以及这些蛋白激酶修饰的伸长体组成蛋白中的特定位点。通过定义磷酸化的单个位点，我们可以产生完全不能磷酸化或模仿磷酸基团存在的突变体。我们将使用这样的突变体来建立一个详细的画面，具体的磷酸化事件如何影响伸长器的功能。

项目成果

Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator.

• DOI: 10.3390/toxins9090272
• 发表时间: 2017-09-05
• 期刊: Toxins
• 影响因子: 4.2
• 作者: Mehlgarten C;Prochaska H;Hammermeister A;Abdel-Fattah W;Wagner M;Krutyhołowa R;Jun SE;Kim GT;Glatt S;Breunig KD;Stark MJR;Schaffrath R
• 通讯作者: Schaffrath R

Insight and Control of Infectious Disease in Global Scenario

全球情景下传染病的洞察与控制

• DOI: 10.5772/31680
• 发表时间: 2012
• 作者: Uthman S

The amidation step of diphthamide biosynthesis in yeast requires DPH6, a gene identified through mining the DPH1-DPH5 interaction network.

• DOI: 10.1371/journal.pgen.1003334
• 发表时间: 2013
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Uthman S;Bär C;Scheidt V;Liu S;ten Have S;Giorgini F;Stark MJ;Schaffrath R
• 通讯作者: Schaffrath R

Decoding the biosynthesis and function of diphthamide, an enigmatic modification of translation elongation factor 2 (EF2).

• DOI: 10.15698/mic2014.06.151
• 发表时间: 2014-05-20
• 期刊: Microbial cell (Graz, Austria)
• 作者: Schaffrath R;Stark MJ
• 通讯作者: Stark MJ