Role of MHV68 v-cyclin in virus egress

MHV68 v-cyclin 在病毒流出中的作用

• 批准号: 8807186
• 负责人: SAMUEL H SPECK
• 金额: $ 23.4万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8807186
• 关键词: 1 year old; 10 year old; Actins; Acute; Adenoviruses; Animal Model; Apoptosis; Attention; B-Lymphocytes; Binding; Biochemical; Biological Models; CDK2 gene; Cell Cycle; Cell Cycle Progression; Cell Death Induction; Cell Line; Cells; Complex; Cyclin A; Cyclins; Cytoskeletal Modeling; Cytoskeleton; DNA Viruses; Defect; Development; Disease; Environment; Epithelial Cells; Exhibits; Foundations; Future; Genes; Growth; HIV; Health; Herpesviridae; Herpesviridae Infections; Homologous Gene; Human Herpesvirus 4; Human Herpesvirus 8; In Vitro; Individual; Infection; Life Cycle Stages; Lung; Lymphocyte Function; Lymphoma; Modeling; Mus; Necrosis; Oncogenes; Papillomavirus; Phenotype; Play; Polyomavirus; Property; Proteins; Rhadinovirus; Role; Saimiriine Herpesvirus 2; Structural Models; Structure; Technetium Tc 99m ciprofloxacin; Transgenes; Transgenic Mice; Viral; Virion; Virus; Virus Diseases; Virus Replication; Work; base; cell type; gamma-2 herpesvirus; gammaherpesvirus; immunosuppressed; in vivo; insight; macrophage; mutant; novel; reactivation from latency; tissue culture; trafficking; tumor; viral cyclin

DESCRIPTION (provided by applicant): Gammaherpesviruses are associated with the development of lympho-proliferative disorders and lymphoma, particularly in immunosuppressed individuals. Indeed, half of the lymphomas that arise in HIV infected individuals are associated with either EBV or KSHV infection. Perturbation of host cell cycle is a common strategy employed by DNA viruses to achieve a cellular environment conducive to viral growth. Adenovirus, polyoma virus, papilloma virus, and many herpesviruses encode genes that directly alter the host cell cycle, or interact with host gene products to the same end. Rhadinoviruses (γ2-herpesviruses), such as Kaposi sarcoma-associated herpesvirus (KSHV), herpesvirus saimiri (HVS), and murine gammaherpesvirus-68 (MHV68), encode a homolog of mammalian D-type cyclins. We have previously shown that the MHV68 v-cyclin is required for: (i) efficient acute replication in the lungs of mice; and (ii) reactivation from latently infected macrophages and B cells. We have recently identified a tissue culture model that recapitulates the replication defect observed with v-cyclin null and v-cyclin CDK binding mutants in vivo. Further characterization of MHV68 replication in this tissue culture model has identified a profound defect in the egress of v-cyclin null and v-cyclin CDK binding mutants from infected cells. In this new R21 application, we propose to investigate the role that the MHV68 v-cyclin plays in virus egress/release from lung epithelial cells. The specific aims are as follows: Aim 1. Characterization of the v-cyclin null mutant MHV68 egress phenotype: 1.a Localization of virions in wt and v-cyclin mutant infected cells; 1.b Cellular localization of v-cyclin during virus infecton; 1.c Assess egress phenotype of a KSHV v-cyclin null mutant in HUVECs. Aim 2. Analysis of v-cyclin functions in the absence of MHV68 infection: 2.a Generate and characterize inducible v-cyclin expressing cell lines; 2.b Identify v-cyclin interacting partners.

描述（由申请方提供）：γ疱疹病毒与淋巴增生性疾病和淋巴瘤的发生相关，特别是在免疫抑制个体中。事实上，在HIV感染者中出现的淋巴瘤中有一半与EBV或KSHV感染有关。干扰宿主细胞周期是DNA病毒获得有利于病毒生长的细胞环境的常用策略。腺病毒、多瘤病毒、乳头瘤病毒和许多疱疹病毒编码直接改变宿主细胞周期或与宿主基因产物相互作用以达到相同目的的基因。杆状病毒（γ2-疱疹病毒），如卡波西肉瘤相关疱疹病毒（KSHV）、松鼠猴疱疹病毒（HVS）和鼠γ疱疹病毒-68（MHV 68），编码哺乳动物D型细胞周期蛋白的同源物。我们以前已经表明，MHV 68 v-细胞周期蛋白是必需的：（i）在小鼠肺中的有效急性复制;和（ii）从潜伏感染的巨噬细胞和B细胞的再活化。我们最近确定了一种组织培养模型，概括了在体内观察到的v-cyclin null和v-cyclin CDK结合突变体的复制缺陷。在该组织培养模型中对MHV 68复制的进一步表征已经确定了v-细胞周期蛋白无效和v-细胞周期蛋白CDK结合突变体从感染细胞中流出的严重缺陷。在这 新的R21应用，我们建议调查的作用，MHV 68 v-细胞周期蛋白在病毒的出口/释放从肺上皮细胞。具体目标如下：目标1。v-细胞周期蛋白无效突变体MHV 68外出表型的表征：1.a病毒体在wt和v-细胞周期蛋白突变体感染的细胞中的定位; 1.b病毒感染期间v-细胞周期蛋白的细胞定位; 1.c评估HUVEC中KSHV v-细胞周期蛋白无效突变体的外出表型。目标二。在不存在MHV 68感染的情况下分析v-细胞周期蛋白功能：2.a产生和表征可诱导的v-细胞周期蛋白表达细胞系; 2.b鉴定v-细胞周期蛋白相互作用伴侣。