Structural basis for the bacterial transcription-repair coupling factor/RNA polymerase interaction
Structural basis for the bacterial transcription-repair coupling factor/RNA polymerase interaction
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DOI：
10.1093/nar/gkq692
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发表时间：
2010-12-01
影响因子：
14.9
通讯作者：
Darst, Seth A.
中科院分区：
文献类型：
--
作者：
Westblade, Lars F.;Campbell, Elizabeth A.;Darst, Seth A.
转录修复偶联因子(TRCF，mfd基因产物)是一种广泛保守的细菌蛋白，介导转录偶联DNA修复。TRCF利用其依赖于ATP的DNA转位酶活性来移除停滞在DNA损伤部位的转录复合体，并通过将核苷酸切除修复途径的组成部分招募到该部位来刺激修复。TRCF与RNA聚合酶β亚基(RNAP)之间的蛋白质/蛋白质相互作用是TRCF功能所必需的。CARD(也称为CDNL)是结核分枝杆菌rRNA转录的重要调节因子，它与TRCF共享一个同源的RNAP相互作用结构域，也与RNAPβ亚基相互作用。我们测定了TRCF的RNAP相互作用结构域与含有TRCF相互作用决定因素的RNAP-β1结构域的2.9-A分辨率X射线晶体结构。该结构揭示了TRCF/RNAP蛋白质/蛋白质界面的细节，为设计和解释探测TRCF的实验提供了基础，并通过同源性卡片、功能和与RNAP的相互作用。
The transcription-repair coupling factor (TRCF, the product of the mfd gene) is a widely conserved bacterial protein that mediates transcription-coupled DNA repair. TRCF uses its ATP-dependent DNA translocase activity to remove transcription complexes stalled at sites of DNA damage, and stimulates repair by recruiting components of the nucleotide excision repair pathway to the site. A protein/protein interaction between TRCF and the beta-subunit of RNA polymerase (RNAP) is essential for TRCF function. CarD (also called CdnL), an essential regulator of rRNA transcription in Mycobacterium tuberculosis, shares a homologous RNAP interacting domain with TRCF and also interacts with the RNAP beta-subunit. We determined the 2.9-A resolution X-ray crystal structure of the RNAP interacting domain of TRCF complexed with the RNAP-beta 1 domain, which harbors the TRCF interaction determinants. The structure reveals details of the TRCF/RNAP protein/protein interface, providing a basis for the design and interpretation of experiments probing TRCF, and by homology CarD, function and interactions with the RNAP.