EBV BZLF1 GENE PRODUCT

EBV BZLF1 基因产品

• 批准号: 2857158
• 负责人: ERIK K FLEMINGTON
• 金额: $ 25.46万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2857158
• 关键词: DNA replication; Epstein Barr virus; cell growth regulation; gene deletion mutation; gene expression; genetic library; genetic regulatory element; genetic transcription; intermolecular interaction; molecular cloning; nucleic acid sequence; protein structure function; site directed mutagenesis; transcription factor; transfection; virus genetics; virus protein; yeast two hybrid system

EBV is the casual agent of infectious mononucleosis and is associated with the development of both B-cell and epithelial cell malignancies including the endemic form of Burkitt's lymphoma, post-transplantation lympho-proliferative diseases, AIDS-associated lymphomas, Hodgkin's disease and undifferentiated nasopharyngeal carcinoma. Viral genes expressed during the latent phase of the EBV life cycle are growth promoting and are responsible for EBV's link to these human cancers. However, it has been known for some time that the lytic phase of the EBV life cycle occurs in differentiated and/or growth arrested tissues. We have recently found that the lytic switch gene, Zta, has potent cell growth inhibitory activity suggesting that EBV can actively promote this cell growth arrested status. Therefore, Zta may represent an evolutionary counterpart to the latency associated EBV gene products. The objectives of this proposal are to identify how Zta integrates into cell-cycle control pathways and to learn how interactions with key cell- cycle control proteins influences progression through the EBV lytic replication cycle. Our specific aims are: 1) Genetic analysis of Zta mediated cellular growth arrest. a) Generation of Zta mutants (rationale) b) Preliminary characterization of Zta mutants - analysis of dimerization/DNA binding, nuclear localization, transactivation. c) Genetic analysis of Zta mediated growth arrest, p21, p27, and p53 induction, and inhibition of c-Myc expression. 2) Yeast two -hybrid for screening Zta interacting factors. a)Library screening. b) Clone selection - rationale. c) Interaction studies. d) Functional analysis of Zta:Zta-targeting factor interactions. 3) Genetic analysis of Zta mediated latency disruption. a) Development of model system. b) Analysis of latency disruption by Zta mutants.

EBV是传染性单核细胞增多症的致病因子， 随着B细胞和上皮细胞恶性肿瘤的发展 包括地方性伯基特淋巴瘤，移植后 淋巴增生性疾病，艾滋病相关淋巴瘤，霍奇金淋巴瘤 疾病和未分化鼻咽癌。 病毒基因 在潜伏期内表达 EB病毒生命周期的阶段是生长 促进并负责EBV与这些人类癌症的联系。 然而，一段时间以来，人们已经知道，EBV的裂解相 生命周期发生在分化的和/或生长停滞的组织中。 我们 最近发现，裂解开关基因Zta具有强大的细胞毒性， 生长抑制活性表明EBV可以积极促进这一点 细胞生长停滞状态。因此，Zta可以表示 与潜伏期相关的EBV基因产物的进化对应物。 本提案的目标是确定Zta如何融入 细胞周期控制途径，并了解如何与关键细胞的相互作用， 周期控制蛋白通过EBV裂解影响进展 复制周期 本研究的具体目的是：1）Zta的遗传分析 介导的细胞生长停滞。 a）Zta突变体的产生 （基本原理）B）Zta突变体的初步表征-分析 二聚化/DNA结合，核定位，反式激活。c）、 Zta介导的生长停滞、p21、p27和p53的遗传分析 诱导和抑制c-Myc表达。 2)酵母双杂交 筛选Zta相互作用因子。1.图书馆筛选。 B）克隆 选择-理由。（c）相互作用研究。D.功能分析 Zta：Zta靶向因子相互作用。 3)Zta的遗传分析 介导的延迟中断。 （1）模型系统的开发。 B） Zta突变体的潜伏期破坏分析。