MOLECULAR BIOLOGY OF TRANSFORMATION BY EPSTEIN-BARR VIRUS

Epstein-Barr 病毒转化的分子生物学

• 批准号: 3768861
• 负责人: J I COHEN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3768861
• 关键词: B lymphocyte; Epstein Barr virus; SCID mouse; cell transformation; clone cells; fusion gene; gene expression; genetic transcription; leukocyte activation /transformation; nucleoproteins; transfection; transforming virus; virus protein

Epstein-Barr virus (EBV) nuclear protein 2 (EBNA-2) is essential for B cell transformation by the virus. The goal of this project is to determine the mechanism of B cell transformation by EBV. Injection of EBV-transformed human lymphocytes into SCID mice results in the appearance of fatal human B cell tumors containing EBV. Cell lines were developed that contained EBV mutants that were identical, except for different deletions in their EBV EBNA-2 gene. EBV mutants with deletions in the amino half of EBNA-2 had their EBV EBNA-2 gene. EBV mutants with deletions in the amino half of EBNA-2 had reduced transforming activity in vitro and produced tumors in mice more slowly, while a carboxy- terminal mutant had a transforming activity greater than or equal to wild-type and produced tumors more rapidly than wild-type. Therefore, EBNA-2 is important both for lymphocyte transformation in cell culture and B cell tumor growth in SCID mice. EBV EBNA-2 contains a domain that directly activates transcription; a 14 amino acid region possesses much of the transcriptional activity and is essential for transformation by EBV. A chimeric virus was constructed that contained a portion of the transcriptional activation domain from the herpes simplex virus VP16 inserted in place of the 14 amino acid region in EBNA-2. This chimeric virus was able to transform B lymphocytes and transactivate expression of EBV and B cell genes. Mutation of a single amino acid in the 14 amino acid domain of EBNA-2 abolished transcriptional activation and B cell transformation. Thus, transcriptional activation is required for EBV-induced B cell transformation.

EB病毒（EBV）核蛋白2（EBNA-2）对B是必需的 细胞被病毒转化。 该项目的目标是 明确EBV对B细胞转化的机制。 将EB病毒转化的人淋巴细胞注射到SCID小鼠中， 含有EBV的致命性人类B细胞肿瘤的出现。 细胞系 开发了包含相同的EBV突变体的病毒，除了 在他们的EBV EBNA-2基因中有不同的缺失。 EBV缺失突变体 在EBNA-2的氨基半部分具有它们的EBV-EBNA-2基因。 EBV突变体， EBNA-2氨基部分的缺失降低了转化活性 在体外和小鼠中产生肿瘤的速度更慢，而羧基- 末端突变体的转化活性大于或等于 野生型，并且比野生型更快地产生肿瘤。 因此，我们认为， EBNA-2对于细胞培养中的淋巴细胞转化和细胞增殖都是重要的。 和B细胞肿瘤生长。 EBV-EBNA-2含有一个直接激活转录的结构域; 氨基酸区域具有大部分的转录活性， 是EBV转化的关键。 构建了嵌合病毒 它含有一部分转录激活结构域， 单纯疱疹病毒VP 16插入14个氨基酸的位置 EBNA-2中的区域。 这种嵌合病毒能够转化B 并反式激活EBV和B细胞基因的表达。 EBNA-2的14个氨基酸结构域中的单个氨基酸突变 消除了转录激活和B细胞转化。 因此，在本发明中， EBV诱导的B细胞需要转录激活 转型