Epstein-Barr virus nuclear protein 3C binds to the N-terminal (NTD) and beta trefoil domains (BTD) of RBP/CSL; Only the NTD interaction is essential for lymphoblastoid cell growth
Epstein-Barr virus nuclear protein 3C binds to the N-terminal (NTD) and beta trefoil domains (BTD) of RBP/CSL; Only the NTD interaction is essential for lymphoblastoid cell growth
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DOI：
10.1016/j.virol.2011.02.018
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发表时间：
2011-05-25
期刊：
影响因子：
3.7
通讯作者：
Johannsen, Eric
中科院分区：
文献类型：
--
作者：
Calderwood, Michael A.;Lee, Sungwook;Johannsen, Eric
EBV核蛋白EBNA2、EBNA3A和EBNA3C与RBP/CSL的关联是淋巴母细胞样细胞系（LCL）增殖所必需的。RBP/CSL相互作用所需的EBNA3同源结构域中的保守残基缺乏介导EBNA2和Notch结合到RBP/CSL β -三叶结构域（BTD）的WIT基序。我们在EBNA3A（aa128-204）和EBNA3C（aa211-233）中绘制了RBP/CSL相互作用残基。EBNA3A的结果与先前的报道（aa125-222）一致，但EBNA3C结构域出乎意料地小，并且包含一个“WTP”序列。这个EBNA3C WTP基序在体外、酵母和哺乳动物细胞中赋予RBP/CSL结合。此外，EBNA3C WTP -> STP（W227S）突变破坏了BID结合，而EBNA3同源结构域突变破坏了RBP/CSL n端结构域（NTD）结合。在报告性实验中，WTP对EBNA3C抑制EBNA2或维持LCL生长并不是必需的。我们的研究结果表明，EBNA3蛋白与多个RBP/CSL结构域相互作用，但LCL生长只需要NTD相互作用。(C) 2011爱思唯尔公司版权所有。
Association of EBV nuclear proteins EBNA2, EBNA3A and EBNA3C with RBP/CSL, is essential for lymphoblastoid cell line (LCL) proliferation. Conserved residues in the EBNA3 homology domain, required for RBP/CSL interaction, lack the WIT motif that mediates EBNA2 and Notch binding to the RBP/CSL beta-trefoil domain (BTD). We map RBP/CSL interacting residues within EBNA3A(aa128-204) and EBNA3C(aa211-233). The EBNA3A results are consistent with an earlier report (aa125-222), but the EBNA3C domain is unexpectedly small and includes a "WTP" sequence. This EBNA3C WTP motif confers RBP/CSL binding in vitro, in yeast, and in mammalian cells. Further, an EBNA3C WTP -> STP(W227S) mutation impaired BID binding whereas EBNA3 homology domain mutations disrupted RBP/CSL N-terminal domain (NTD) binding. WTP was not essential for EBNA3C repression of EBNA2 in reporter assays or for maintenance of LCL growth. Our results indicate that EBNA3 proteins interact with multiple RBP/CSL domains, but only NTD interactions are required for LCL growth. (C) 2011 Elsevier Inc. All rights reserved.