EBNA-3C in B Lymphocyte Transformation by EBV

EBNA-3C 在 EBV 转化 B 淋巴细胞中的作用

• 批准号: 6532875
• 负责人: ERIC C JOHANNSEN
• 金额: $ 12.85万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6532875
• 关键词: B lymphocyte; Epstein Barr virus; chimeric proteins; clinical research; human subject; neoplastic transformation; protein structure function; recombinant virus; tissue /cell culture; virus antigen

DESCRIPTION (provided by applicant): This project will attempt to define the mechanism(s) by which the Epstein-Barr virus nuclear antigen 3C (EBNA-3C) promotes B lymphocyte transformation into lymphoblastoid cell lines (LCLs). Previous studies have established that EBV genomes lacking an intact EBNA-3C cannot growth transform B cells. The central hypothesis of this application is that EBNA-3C is essential for the establishment and maintenance of LCLs and that it exerts its effect by regulating the expression of critical cellular and viral genes. The requirement for continued EBNA-3C expression in the maintenance of LCLs will be tested using a recombinant EBV expressing a hydroxytamoxifen/EBNA-3C fusion protein. It is anticipated that, in the absence of hydroxytamoxifen, the tethering of this fusion protein in the cytoplasm will lead to growth arrest and/or apoptosis. In this case, the ability of transfected EBNA-3C as well as multiple EBNA-3C mutants to complement this defect will be assessed. If instead, EBNA-3C proves to be dispensable for the maintenance of transformation, a genetic analysis of EBNA- 3C mutants will be conducted for their ability to mediate the establishment of LCLs. The ability of these mutants to synergistically co-activate with the major viral transactivator (EBNA-2) will also be assessed. Using the above information, the transcriptional profile of EBNA-3C transformation competent mutants will be contrasted to mutants unable to mediate transformation in an effort to establish the major pathways through which EBNA-3C promotes B lymphocyte transformation. Biochemical methods including a yeast two hybrid screen and co-immunoprecipitation will also be employed to discover what cellular proteins interact with the identified EBNA-3C effector domains.

描述（由申请人提供）：该项目将尝试定义Epstein-Barr病毒核抗原3C（EBNA-3C）的机制 促进B淋巴细胞转化为淋巴细胞细胞系（LCLS）。 先前的研究表明，EBV基因组缺乏完整的EBNA-3C 无法生长转化B细胞。该应用的中心假设是 EBNA-3C对于建立和维护LCL和 它通过调节临界细胞的表达来发挥其作用 和病毒基因。在持续EBNA-3C表达中的要求 LCL的维护将使用表达的重组EBV测试 羟基昔芬/EBNA-3C融合蛋白。可以预料，在 缺乏羟基莫昔芬，这种融合蛋白在 细胞质将导致生长停滞和/或凋亡。在这种情况下， 转染的EBNA-3C以及多个EBNA-3C突变体的能力 补充该缺陷将被评估。如果相反，EBNA-3C被证明是 可用于维持转化的可分配，EBNA-的遗传分析 将进行3C突变体，以调解建立的能力 LCLS。这些突变体协同激活与 还将评估主要的病毒反式激活因子（EBNA-2）。使用以上 信息，EBNA-3C转换的转录曲线胜任 突变体将与无法介导的突变体形成鲜明对比 努力建立EBNA-3C促进B的主要途径 淋巴细胞转化。生化方法，包括酵母两杂种 还将采用屏幕和共免疫沉淀来发现什么 细胞蛋白与已鉴定的EBNA-3C效应子结构域相互作用。