INTERFERON REGULATORY FACTORS CELL CYCLE AND EBV LATENCY

干扰素调节因子细胞周期和 EBV 潜伏期

• 批准号: 6328770
• 负责人: JOSEPH S PAGANO
• 金额: $ 24.1万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6328770
• 关键词: DNA footprinting; Epstein Barr virus; RNase protection assay; SDS polyacrylamide gel electrophoresis; cell cycle; flow cytometry; gel mobility shift assay; gene expression; gene induction /repression; genetic transcription; host organism interaction; immunoprecipitation; interferon alpha; interferons; laboratory rabbit; latent virus infection; messenger RNA; northern blottings; polymerase chain reaction; protein binding; radiotracer; tissue /cell culture; transcription factor; virus infection mechanism; western blottings

DESCRIPTION: In healthy seropositive persons (and in Burkitt s lymphoma) EBV infection of circulating lymphocytes is in a highly restricted form called type I latency in which only the BamHI Q promoter (Qp) is active, and the EB nuclear antigen 1 (EBNA-1) is expressed. During studies of Qp, which have the long-term objective of understanding how latency is established and maintained and how episomal replication is synchronized with cell cycle, the applicant has isolated a new gene in the family of interferon regulatory factors (IRFs), which are transcriptional factors that bind to the conserved interferon-stimulated responsive element (ISRE) and serve as transactivators or repressors. The protein products of the new gene, IRF-4A, -4B, -4C, bind to an ISRE site in Qp, apparently the first functional ISRE characterized in a viral promoter. Moreover, interferon alpha (IFN-a) appears to activate Qp during primary infection. These findings have opened a new field for exploration, namely, how IRFs and IFN may affect EBV infection, particularly latency. A second mechanism governing Qp centers on regulation of transcription of EBNA1 mRNA by the cell cycle, reciprocally mediated by E2F and EBNA-1. Experiments will address: 1) further characterization of IRF-4 and involvement of other IRF members in Qp regulation; 2) the function of IFN-a in the activation of Qp and its possible biological consequence; 3) timing of expression of EBNA-1 mRNA and protein during cell cycle and mechanism of cell-cycle activation of Qp. Results from the proposed experiments should offer new insights into how these cellular and viral factors unite to regulate transcription of the EBNA-1 gene from Qp, which is central to understanding EBV-host interaction and the viral latency and transformation processes.

描述：在健康的血清阳性人中（在伯基特的淋巴瘤中） 循环淋巴细胞的EBV感染为高度限制的形式 称为I型潜伏期，其中仅BAMHI Q启动子（QP）处于活动状态，并且 表达EB核抗原1（EBNA-1）。 在QP研究中 具有长期目标，以了解如何确定潜伏期和 维护以及如何与细胞周期同步偶发复制 申请人在干扰素调节的家族中分离了一个新基因 因素（IRF），是与保守的转录因子 干扰素刺激的响应元件（ISRE）并用作反式激活因子 或阻遏物。 新基因IRF -4A，-4B，-4C的蛋白质产物结合 到QP中的ISRE站点，显然是第一个功能性的ISRE 病毒启动子。 此外，干扰素α（IFN-A）似乎激活QP 在初次感染期间。 这些发现为 探索，即IRF和IFN如何影响EBV感染，尤其是 潜伏期。 关于QP的第二种机制，以调节 通过细胞周期对EBNA1 mRNA的转录，由E2F相互介导 和EBNA-1。 实验将解决：1）IRF-4的进一步表征 以及其他IRF成员参与QP法规； 2）功能 IFN-A激活QP及其可能的生物学后果； 3） EBNA-1 mRNA和蛋白质表达的时间在细胞周期和 QP的细胞周期激活机理。提议的结果 实验应提供有关这些细胞和病毒的新见解 从QP调节EBNA-1基因转录的因素，这是 了解EBV-host互动和病毒潜伏期的中心 转换过程。