REGULATION OF REPLICATION AND LATENCY BY EBV EBNA-1

爱泼斯坦-巴尔病毒(EBV)引起传染性单核细胞增多症，以 80%的美国成年人口的潜伏感染。个人 免疫功能受损的人，如艾滋病患者和患者 正在接受器官或骨髓移植的人，经常发生 再次激活EBV感染的症状。EBV阳性的发生率 这些患者的淋巴瘤也是一个令人担忧的原因。一个 了解与病毒复制和维持有关的因素 从长远来看，延迟将有助于管理这些 病人。EBV也被认为是非洲Burkitt的一个致病因素 淋巴瘤和鼻咽癌(主要发生在中国人 种族)，因为这些肿瘤的细胞含有EBV DNA并表达 EBNA-1抗原。拟议研究的长期目标是 明确参与Epstein-Barr复制的分子机制 病毒DNA和病毒在转化细胞中潜伏期的维持。 这项应用的具体目标是研究生物和 涉及EBV核抗原EBNA-1和EBV的生化过程 与之相互作用的EBV DNA区域包括：1.分离 完整的EBNA-1蛋白。2.检查装订参数 EBNA-1与其DNA结合位点的结合。3.对生物的检验 操纵EBNA-1的数量和亲和力的意义 ORI-P中的结合位点。4.EBNA-1最小区域的测定 特异性DNA结合、质粒维持、核所需 EBNA-1的定位及其染色体关联。5. EBNA-1在双基因调控中可能作用的研究 表达，以及6.裂解EBV复制组织的鉴定。 特定的试剂将使用重组DNA技术和 检测程序将包括DNAaseI足迹、硝酸纤维素 滤过结合试验、DNA转染法、过氧化氢酶试验和免疫荧光法。

Epstein-Barr Virus (EBV) causes infectious mononucleosis and is carried as a latent infection by 80% of the adult American population. Individuals who are immunologically compromised, such as AIDS victims, and patient undergoing organ or bone-marrow transplantation, frequently develop symptoms of re-activated EBV infection. Incidences of EBV-positive lymphomas in these patients has also been a cause for concern. An understanding of the factors involved in viral replication and maintance of latency would, in the long term, assist in the management of these patients. EBV is also believed to be a causative factor in Africa Burkitt lymphoma and nasopharyngeal carcinoma (which occurs pedominatly in Chinese ethnic groups) since the cells of these tumors contain EBV DNA and express the EBNA-1 antigen. The long range goals fo the proposed research is to define the molecular mechanisms involved in the replication of Epstein-Barr Virus DNA and in the maintenance of viral latency in transformed cells. The specific aims of this application are to investigate the biological and biochemical processes involving the EBV nuclear antigen EBNA-1 and the regions of EBV DNA with which it interacts and will include: 1. Isolation of the intact EBNA-1 protein. 2. Examination of the parameters of binding of EBNA-1 to its DNA binding sites. 3. Examination of the biological implications of manipulating the numbers and affinities of the EBNA-1 binding sites in ori-P. 4. Determination of the minimal region of EBNA-1 required for specific DNA binding, plasmid maintenance, nuclear localization of EBNA-1, and chromosomal association of EBNA-1. 5. Invesitgation of a possible role for EBNA-1 in regulation of biral gene expression, and 6. Identification of lytic EBV orgins of replication. Specific reagents will be generated using recombinant Dna thechnology and assay procedures will include DNAaseI footprinting, nitrocellulose filterbinding assays, DNA transfection, CAT assays and immunoflorescene.