EBNA LP AND EBV INDUCED TRANSFORMATION

EBNA LP 和 EBV 引发的转型

• 批准号: 2057015
• 负责人: JOAN B MANNICK
• 金额: $ 9.15万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-30 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2057015
• 关键词: B lymphocyte; Burkitt's lymphoma; Epstein Barr virus; RNA biosynthesis; RNA splicing; biological models; carcinoma; gene expression; genetic manipulation; immunosuppression; latent virus infection; mutant; nasopharyngeal neoplasms; neoplastic growth; nucleoproteins; protein structure function; sarcoma; viral carcinogenesis; virus genetics; virus replication

Epstein-Barr virus (EBV) is a human herpes virus associated with a variety of tumors including African Burkitt lymphoma, nasopharyngeal carcinoma and immunoblastic sarcoma in immunosuppressed patients. EBV usually establishes a latent infection in B lymphocytes and stimulates them to proliferate. Sustained polyclonal proliferation of EBV infected cells in vivo may occasionally result in monoclonal malignancies. In vitro, B cells are immortalized and provide a model with which to study EBV-induced tumorigenicity. Although the EBV genome contains about 100 genes, only 10 of these are known to be expressed in latently infected, growth transformed lymphocytes. These genes are of interest because they are likely to be involved in the immortalization of lymphocytes as well as in the maintenance of viral latency. One of the genes is the EBV nuclear antigen leader protein (EBNA-LP). The function of EBNA-LP is unknown although preliminary experiments have shown that deletion of the last two exons of EBNA-LP results in a marked reduction in the growth rate of transformed cells. The aim of the proposed studies is to determine the function of EBNA-LP. Recombinant viruses will be constructed with mutations in EBNA-LP using techniques which our laboratory has used successfully to construct recombinant EBV with mutations in the EBNA2 gene. We then will determine if the mutations in EBNA-LP alter growth rate, RNA metabolism or protein expression of cells transformed by EBV. Finally we will analyze whether transfection of wild-type EBNA-LP into cells expressing mutant EBNA-LP can recreate a wild type phenotype, thus confirming that alterations in phenotype are due to the EBNA-LP mutations. Elucidation of the function of EBNA-LP will lead to a better understanding of EBV-induced transformation which in turn may increase our understanding of oncogenesis.

Epstein-Barr 病毒 (EBV) 是一种与多种病毒相关的人类疱疹病毒。 包括非洲伯基特淋巴瘤、鼻咽癌等肿瘤 免疫抑制患者的免疫母细胞肉瘤。 EBV 通常建立 B 淋巴细胞中的潜伏感染并刺激它们增殖。 EBV感染细胞在体内的持续多克隆增殖可能 有时会导致单克隆恶性肿瘤。在体外，B 细胞是 永生化并提供了研究 EBV 诱导的模型 致瘤性。 尽管 EBV 基因组包含大约 100 个基因，但其中只有 10 个是 已知在潜伏感染的生长转化淋巴细胞中表达。 这些基因很有趣，因为它们可能参与 淋巴细胞的永生化以及病毒的维持 延迟。其中基因之一是 EBV 核抗原前导蛋白 （EBNA-LP）。 EBNA-LP 的功能虽然初步了解，但尚不清楚 实验表明，EBNA-LP最后两个外显子的缺失 导致转化细胞的生长速度显着降低。这 拟议研究的目的是确定 EBNA-LP 的功能。 将使用 EBNA-LP 中的突变构建重组病毒 我们实验室已成功使用的技术构建 EBNA2 基因突变的重组 EBV。然后我们将确定是否 EBNA-LP 中的突变会改变生长速度、RNA 代谢或蛋白质 EBV转化细胞的表达。最后我们来分析是否 将野生型 EBNA-LP 转染到表达突变型 EBNA-LP 的细胞中可以 重建野生型表型，从而确认改变 表型是由于 EBNA-LP 突变造成的。功能说明 EBNA-LP 将有助于更好地理解 EBV 诱导的转化 这反过来可能会增加我们对肿瘤发生的理解。