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: 6618097
• 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 2004-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6618097
• 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）：本项目将尝试定义 EB病毒核抗原3C（EBNA-3C） 促进B淋巴细胞转化为类淋巴母细胞系（LCL）。 以前的研究已经确定，缺乏完整EBNA-3C的EBV基因组 不能生长转化B细胞。本申请的中心假设是 EBNA-3C对于建立和维护LCL至关重要， 它通过调节关键细胞因子的表达来发挥作用， 和病毒基因。EBNA-3C在肿瘤细胞中持续表达的需要是一个重要因素。 LCL的维持将使用表达LCL的重组EBV进行测试。 羟基他莫昔芬/EBNA-3C融合蛋白。预计，在 在不存在羟基他莫昔芬的情况下，这种融合蛋白在细胞中的束缚作用是可能的。 细胞质将导致生长停滞和/或细胞凋亡。本案中 转染的EBNA-3C以及多种EBNA-3C突变体的能力， 将对这一缺陷进行评估。如果相反，EBNA-3C被证明是 为了维持转化，EBNA的遗传分析- 3C突变体将进行其介导的能力，建立 LCL。这些突变体协同协同激活的能力， 还将评估主要病毒反式激活因子（EBNA-2）。使用上述 信息，EBNA-3C转化能力的转录谱 将突变体与不能介导转化的突变体进行对比， 努力建立EBNA-3C促进B的主要途径 淋巴细胞转化包括酵母双杂交在内的生化方法 筛选和免疫共沉淀也将被用来发现 细胞蛋白与鉴定的EBNA-3C效应域相互作用。