Dynamic Protein-Protein Interaction Wiring of the Human Spliceosome
Dynamic Protein-Protein Interaction Wiring of the Human Spliceosome
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DOI：
10.1016/j.molcel.2011.12.034
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发表时间：
2012-02-24
期刊：
影响因子：
16
通讯作者：
Stelzl, Ulrich
中科院分区：
文献类型：
--
作者：
Hegele, Anna;Kamburov, Atanas;Stelzl, Ulrich
超过200种蛋白质与剪接体共纯化，剪接体是催化前体mRNA剪接的组成动态RNP。为了更好地了解蛋白质-蛋白质相互作用的剪接，我们系统地研究了人类剪接体蛋白之间的相互作用。一个全面的Y2 H相互作用矩阵筛选产生了蛋白质相互作用图，包括196个蛋白质之间的632个相互作用。其中，242相互作用之间的剪接体核心蛋白，并在很大程度上验证了免疫共沉淀。为了揭示蛋白质相互作用的动态变化，我们将剪接体复合物纯化信息与我们的相互作用数据相结合，并进行链接聚类。这些数据以及相互作用竞争实验表明，在剪接的第1步中，hPRP 8与SF 3b蛋白的相互作用被hSLU 7取代，将该第二步因子定位在活性位点附近，并且DEAH盒解旋酶hPRP 2和hPRP 16通过与GPKOW的有序相互作用进行合作。我们的数据提供了广泛的信息spliceosomal蛋白质相互作用网络及其动力学。
More than 200 proteins copurify with spliceosomes, the compositionally dynamic RNPs catalyzing pre-mRNA splicing. To better understand protein protein interactions governing splicing, we systematically investigated interactions between human spliceosomal proteins. A comprehensive Y2H interaction matrix screen generated a protein interaction map comprising 632 interactions between 196 proteins. Among these, 242 interactions were found between spliceosomal core proteins and largely validated by coimmunoprecipitation. To reveal dynamic changes in protein interactions, we integrated spliceosomal complex purification information with our interaction data and performed link clustering. These data, together with interaction competition experiments, suggest that during step 1 of splicing, hPRP8 interactions with SF3b proteins are replaced by hSLU7, positioning this second step factor close to the active site, and that the DEAH-box helicases hPRP2 and hPRP16 cooperate through ordered interactions with GPKOW. Our data provide extensive information about the spliceosomal protein interaction network and its dynamics.