CONTROL OF EBV LYTIC GENE EXPRESSION DURING LATENCY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。本研究的主要目的是阐明EB病毒(EBV)感染的B淋巴细胞从潜伏复制到病毒复制的控制机制。更具体地说，这项研究集中在两个连锁的EBV基因BSLF1和BRLF1的调控上，这两个基因与这种开关密切相关。详细了解病毒重新激活是如何控制的，对于理解潜伏期的维持是至关重要的，并可能揭示干扰病毒在宿主中持续的策略。在过去的资助期间，在确定调控BZLF1基因启动子(ZP)的关键顺式元件以及与这些位点结合的细胞因子方面取得了重大进展。此外，还发现了两条钙反应通路，它们可以触发病毒重新激活。然而，我们对BZLF1基因的转录是如何调控的只有部分了解。此外，BRLF1基因的调控还没有得到仔细的分析。因此，为了鉴定和进一步确定参与调节BRLF1/BZLF1基因座的顺式元件、细胞转录因子和信号通路，我们提出了以下目标：目的1.调节ZP和Rp的顺式元件的功能分析：1A。研究BRLF1和BZLF1基因之间的相互依赖关系；鉴定和鉴定参与调节RP诱导的顺式元件；1c。鉴定SP1、SP3和MEF2D结合Z1结构域在ZP；1D中的作用。鉴定和克隆与ZP中负顺式元件ZIIR结合的细胞因子(S)；目的2.BRLF1/BZLF1基因座EB病毒突变体的产生：2A。表征ZP和Zta在诱导病毒裂解周期中的作用；评估参与调节ZP和RP的关键顺式元件中选定突变的影响。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The major goal of this research is the elucidation of the mechanisms controlling the switch from latency to viral replication in Epstein-Barr virus (EBV) infected B lymphocytes. More specifically, this research focuses on the regulation of two linked EBV genes, BSLF1 and BRLF1, which are integrally involved in this switch. A detailed understanding of how viral reactivation is controlled is essential for understanding maintenance of latency, and may reveal strategies for interfering with viral persistence in the host. During the pervious funding period, significant progress has been made in identifying the critical cis-elements involved in regulating the BZLF1 gene promoter (Zp), and the cellular factors that bind to these sites. In addition, two calcium response pathways have been identified that can trigger viral reactivation. However, we only have a partial picture of how transcription of the BZLF1 gene is regulated. Furthermore, regulation of the BRLF1 gene has not been carefully analyzed. Thus, to identify and further define the cis-elements, cellular transcription factors and signaling pathways involved regulating the BRLF1/BZLF1 gene locus we propose the following aims: Aim 1. Functional analysis of cis-elements involved in regulating ZP and RP: 1a. Investigate the interdependence of the BRLF1 and BZLF1 genes; 1b. Identify and characterize cis-elements involved in regulating induction of Rp; 1c. Characterize role of Sp1, Sp3 and MEF2D binding to the Z1 domains in Zp; 1d. Characterize and clone the cellular factor(s) binging to the negative cis-element ZIIR in Zp; Aim 2. Generation of EBV mutants in the BRLF1/BZLF1 locus: 2a. Characterize the role of Zp and Zta in the induction of the viral lytic cycle; 2b. Assess the impact of selected mutations in critical cis-elements involved in regulating Zp and Rp.