Characterization of the gamma HV68 v-cyclin

描述（由申请人提供）：这是第一次更新的授权，以调查细胞周期蛋白同系物编码的机制？-疱疹病毒（v-细胞周期蛋白）有助于？疱疹病毒发病机制和潜伏期。人类？疱疹病毒KSHV和EBV是癌症的重要原因，特别是在免疫功能低下的个体中。由于这些病毒的种属特异性，其发病机制的体内研究受到限制。我们和其他人一直在开发一种小动物模型系统，用小鼠γ疱疹病毒68（yHV 68）感染近交系小鼠，用于分析？疱疹病毒感染这个应用程序的重点是作用的？HV 68 v-cyclin在疾病发病机制中的作用。值得注意的是，？2-疱疹病毒（HVS、KSHV和yHV 68）都编码D型细胞周期蛋白的同源物，而EBV感染上调宿主D型细胞周期蛋白的表达。因此，我们希望分析的？HV 68 v-cyclin将为保守的致病机制提供重要的见解。我们已经表明，？HV 68 v-细胞周期蛋白是一个癌基因，促进细胞周期的进展，在初级淋巴细胞和a？HV 68 v-细胞周期蛋白突变体重新激活无效从潜伏感染M？和/或B细胞（进展报告）。这些观察得出以下4个目标。 目标1。v-细胞周期蛋白在慢性感染期间的作用[评估剂量和感染途径对急性病毒复制、潜伏期和再活化的影响; wt和v-细胞周期蛋白无效yHV 68的体内感染竞争分析;进一步表征v-细胞周期蛋白无效yHV 68再活化缺陷;表征携带KSHV v-细胞周期蛋白的嵌合病毒]。 目标二。v-细胞周期蛋白基因表达的转录调控[裂解周期相关v-细胞周期蛋白基因转录的作用;潜伏/再活化相关剪接v-细胞周期蛋白基因转录物的分析; P1和P2启动的拉娜/v-细胞周期蛋白剪接转录物的表征]。 目标3：开发和表征组织培养模型，以研究v-细胞周期蛋白功能[急性病毒复制期间依赖于CDK的功能;病毒从潜伏期再激活期间不依赖于CDK的功能]。 目标4。表征不依赖于CDK的v-细胞周期蛋白功能[绘制病毒再活化所需的结构域;比较野生型和CDK结合突变型v-细胞周期蛋白的基因表达谱;鉴定和表征与v-细胞周期蛋白相互作用的细胞和病毒蛋白]。

DESCRIPTION (provided by applicant): This is the first renewal of a grant to investigate the mechanisms by which cyclin homologs encoded by ?-herpesviruses (v-cyclins) contribute to ?-herpesvirus pathogenesis and latency. The human ?-herpes viruses KSHV and EBV are important causes of cancer, especially in immunocompromised individuals. Because of the species specificity of these viruses, in vivo studies of their pathogenesis have been limited. We and others have been developing a small animal model system, infection of inbred mice with murine gammaherpesvirus 68 (yHV68), for analysis of the pathogenesis of ?-herpesvirus infection. This application focuses on the role of the ?HV68 v-cyclin in disease pathogenesis. Notably, the ?2-herpesviruses (HVS, KSHV and yHV68) all encode homologs of D-type cyclins, while EBV infection upregulates expression of host D-type cyclins. Thus, we expect analysis of the ?HV68 v-cyclin will provide important insights into a conserved pathogenic mechanism. We have shown that the ?HV68 v-cyclin is an oncogene that promotes cell cycle progression in primary lymphocytes and that a ?HV68 v-cyclin mutant reactivates inefficiently from latently infected M? and/or B cells (Progress Report). These observations lead to the following 4 aims. Aim 1. Role of v-cyclin during chronic infection [assess impact of dose and route of infection on acute virus replication, latency and reactivation; in vivo infection competition analysis with wt and v-cyclin null yHV68; further characterize v-cyclin null yHV68 reactivation defect; characterization of chimeric virus harboring KSHV v-cyclin]. Aim 2. Transcriptional regulation of v-cyclin gene expression [role of lytic cycle-associated v-cyclin gene transcription; analysis of latency/reactivation-associated spliced v-cyclin gene transcripts; characterization of P1- and P2-initiated LANA/v-cyclin spliced transcripts]. Aim 3. Develop and characterize tissue culture models to investigate v-cyclin functions [CDK-dependent function during acute virus replication; CDK-independent function during virus reactivation from latency]. Aim 4. Characterize CDK-independent v-cyclin function [map domains required for virus reactivation; gene expression profiling comparing wt and CDK-binding mutant v-cyclin; identify and characterize cellular and viral proteins interacting with v-cyclin].