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Author:
Date:
2013-09-20
[Abstract] Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA2) induces expression of both viral and cellular genes in virus infected B cells by mimicking activated Notch receptors (Notch-IC) that mediate transcription activation through binding to the repressing domain of the recombining binding protein suppressor of hairless (RBP-Jκ). In general, chromatin immunoprecipitation (ChIP) assays, electrophoresis mobility shift assays (EMSA), streptavidin-agarose mediated DNA pull-down assays, together with cell-based transcription reporter assays were conducted to verify whether the query protein is involved in EBNA2-dependent transcription. The ATP-bound state of nuclear chaperone nucleophosmin (NPM1) has been implicated in pleiotropic biological processes. An ATP-agarose-mediated pull-down protocol was ...
[摘要] EB病毒（EBV）核抗原2（EBNA2）通过模拟激活的Notch受体（Notch-IC）诱导病毒感染的B细胞中病毒和细胞基因的表达，所述Notch受体通过结合重组结合的抑制结构域介导转录激活无毛蛋白抑制剂（RBP-Jκ）。通常，进行染色质免疫沉淀（ChIP）测定，电泳迁移率变动测定（EMSA），链霉亲和素 - 琼脂糖介导的DNA下拉测定以及基于细胞的转录报道基因测定，以验证查询蛋白是否参与EBNA2依赖性转录。核伴侣核蛋白（NPM1）的ATP结合状态已涉及多效生物过程。开发ATP-琼脂糖介导的下拉方案以监测由ATP结合的NPM1诱导的引发前复合物的形成。根据EBNA2和Notch-IC已经显示出在B细胞系中靶基因的活化方面是部分可互换的，可以想象EBNA2是活化的Notch IC的生物学等价物。
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