High-resolution mapping of the protein interaction network for the human transcription machinery and affinity purification of RNA polymerase II-associated complexes
High-resolution mapping of the protein interaction network for the human transcription machinery and affinity purification of RNA polymerase II-associated complexes
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DOI：
10.1016/j.ymeth.2009.05.005
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发表时间：
2009-08-01
期刊：
影响因子：
4.8
通讯作者：
Coulombe, Benoit
中科院分区：
文献类型：
--
作者：
Cloutier, Philippe;Al-Khoury, Racha;Coulombe, Benoit
30年来对基因转录的研究已经揭示了在mRNA合成期间直接或间接调节RNA聚合酶II（RNAPII）活性的无数因素。然而，许多调节因素仍有待发现。使用蛋白质亲和纯化耦合质谱（AP-MS），我们最近解开了RNAPII和它的辅助因子从人细胞提取物的可溶性部分形成的高密度相互作用网络。使用经过训练以最大限度地降低假阳性和假阴性率的机器学习方法对数据集进行验证，产生了高置信度的数据集，并发现了调节RNAPII转录机制的新型相互作用物，包括我们命名为RNAPII相关蛋白3（RPAP 3）复合物的新蛋白质组装。(C)2009 Elsevier Inc. All rights reserved.
Thirty years of research on gene transcription has uncovered a myriad of factors that regulate, directly or indirectly, the activity of RNA polymerase II (RNAPII) during mRNA synthesis. Yet many regulatory factors remain to be discovered. Using protein affinity purification coupled to mass spectrometry (AP-MS), we recently unraveled a high-density interaction network formed by RNAPII and its accessory factors from the soluble fraction of human cell extracts. Validation of the dataset using a machine learning approach trained to minimize the rate of false positives and false negatives yielded a high-confidence dataset and uncovered novel interactors that regulate the RNAPII transcription machinery, including a new protein assembly we named the RNAPII-Associated Protein 3 (RPAP3) complex. (C) 2009 Elsevier Inc. All rights reserved.