REGULATION OF REPLICATION AND LATENCY BY EBV EBNA-1

EB病毒（EBV）引起传染性单核细胞增多症， 美国80%的成年人都有潜伏感染 个人 免疫功能低下的人，如艾滋病患者和 接受器官或骨髓移植，经常发展 EB病毒感染的症状 EBV阳性的发生率 这些患者中的淋巴瘤也引起了关注。 一个 了解病毒复制和维持的相关因素， 从长远来看，延迟将有助于管理这些 患者 EB病毒也被认为是一个致病因素在非洲伯基特 淋巴瘤和鼻咽癌（主要发生在中国 由于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 定义参与EB病毒复制的分子机制 病毒DNA和维持转化细胞中的病毒潜伏期。 本申请的具体目的是研究生物学和 涉及EBV核抗原EBNA-1和EBNA-2的生物化学过程。 与之相互作用的EBV DNA区域，包括：1.隔离 完整的EBNA-1蛋白。 2.结合参数的检查 EBNA-1的DNA结合位点。 3.生物学检查 操纵EBNA-1的数量和亲和力的影响 ori-P的结合位点。EBNA-1最小区域的确定 所需的特异性DNA结合，质粒维持，核 EBNA-1的定位和EBNA-1的染色体关联。 5. EBNA-1在biral基因调控中的可能作用 表达，6。裂解性EBV复制源的鉴定。 将使用重组DNA技术生产特异性试剂， 测定程序将包括DNA酶I足迹法、硝酸纤维素法 滤膜结合试验、DNA转染、CAT试验和免疫荧光。

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.