EBV BZLF1 GENE PRODUCT

EB病毒是传染性单核细胞增多症的临时病原体，与 随着B细胞和上皮细胞恶性肿瘤的发展 包括移植后的地方性Burkitt淋巴瘤 淋巴增殖性疾病、艾滋病相关淋巴瘤、何杰金氏病 疾病和未分化鼻咽癌。病毒基因 在EBV生命周期的潜伏期表现为生长 促进并对EBV与这些人类癌症的联系负责。 然而，人们已经知道EBV的裂解期 生命周期发生在分化和/或生长受阻的组织中。我们 最近发现裂解开关基因Zta具有强大的细胞 生长抑制活性提示EBV对此有积极的促进作用 细胞生长受阻状态。因此，Zta可能代表一种 与潜伏期相关的EBV基因产物的进化对应。 该提案的目标是确定ZTA如何融入 细胞周期控制途径，并了解如何与关键细胞相互作用- 周期控制蛋白影响EBV裂解的进展 复制周期。我们的具体目标是：1)Zta的遗传分析 导致细胞生长受阻。A)Zta突变体的产生 (原理)b)Zta突变体的初步特征--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基因分析 诱导和抑制c-Myc的表达。2)酵母双杂交 筛选Zta相互作用因素。A)图书馆放映。B)克隆 选择的基本原理。C)互动研究。D)功能分析 Zta：Zta-靶向因子相互作用。3)Zta的遗传分析 调解的延迟中断。A)模型系统的开发。b) Zta突变体的潜伏期干扰分析。

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.