A high resolution protein interaction map of the yeast Mediator complex
A high resolution protein interaction map of the yeast Mediator complex
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DOI：
10.1093/nar/gkh878
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Werner, M
中科院分区：
文献类型：
--
作者：
Guglielmi, B;van Berkum, NL;Werner, M
介体是一种从酵母到人类远程保守的大型模块化蛋白质复合物，其将调节信号从DNA结合转录因子传递到RNA聚合酶II。在酿酒酵母中，Mediator被认为是由24个亚基组成的四个亚复合体，称为头部，中部，尾部和Cdk 8（Srb 8 -11）模块。在这项工作中，我们已经使用筛选和成对双杂交方法来研究芽殖酵母介体亚基之间的蛋白质-蛋白质接触。衍生的相互作用图包括介体亚基之间的许多相互作用域的划定，经常对应于已在进化中保守的片段，以及Cdk 8（Srb 8 -11）和头部模块，头部和中间模块，以及中间和尾部模块之间的新连接。双杂交分析，连同免疫共沉淀研究和凝胶过滤实验显示，Med 31（Soh 1）与酵母介体相关，因此包含25个亚基。最后，对果蝇介体中间模块内的蛋白质相互作用网络的分析表明，从酵母到后生动物，介体复合物的结构组织是保守的。由此产生的相互作用地图提供了一个框架，描绘中介结构功能和调查如何调节中介功能。
Mediator is a large, modular protein complex remotely conserved from yeast to man that conveys regulatory signals from DNA-binding transcription factors to RNA polymerase II. In Saccharomyces cerevisiae, Mediator is thought to be composed of 24 subunits organized in four sub-complexes, termed the head, middle, tail and Cdk8 (Srb8-11) modules. In this work, we have used screening and pair-wise two-hybrid approaches to investigate protein-protein contacts between budding yeast Mediator subunits. The derived interaction map includes the delineation of numerous interaction domains between Mediator subunits, frequently corresponding to segments that have been conserved in evolution, as well as novel connections between the Cdk8 (Srb8-11) and head modules, the head and middle modules, and the middle and tail modules. The two-hybrid analysis, together with co-immunoprecipitation studies and gel filtration experiments revealed that Med31 (Soh1) is associated with the yeast Mediator that therefore comprises 25 subunits. Finally, analysis of the protein interaction network within the Drosophila Mediator middle module indicated that the structural organization of the Mediator complex is conserved from yeast to metazoans. The resulting interaction map provides a framework for delineating Mediator structure-function and investigating how Mediator function is regulated.