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(申请人提供)：伽马疱疹病毒与淋巴增殖性疾病和淋巴瘤的发展有关，特别是在免疫抑制的个人中。事实上，在艾滋病毒感染者中出现的淋巴瘤中，有一半与EBV或KSHV感染有关。扰乱宿主细胞周期是DNA病毒为了获得有利于病毒生长的细胞环境而采取的一种常见策略。腺病毒、多瘤病毒、乳头状瘤病毒和许多疱疹病毒编码的基因可以直接改变宿主细胞周期，或者与宿主基因产物相互作用。卡波西肉瘤相关疱疹病毒(KSHV)、疱疹病毒塞米里病毒(HV)和小鼠伽马疱疹病毒68(MHV68)等横纹状病毒(γ2-疱疹病毒)编码哺乳动物D型细胞周期蛋白的同源物。我们之前已经证明，MHV68v-Cyclin是：(I)在小鼠肺内有效的急性复制；以及(Ii)从潜伏感染的巨噬细胞和B细胞重新激活所必需的。我们最近已经确定了一种组织培养模型，该模型概括了在体内观察到的v-细胞周期蛋白缺失和v-细胞周期蛋白CDK结合突变的复制缺陷。在这个组织培养模型中对MHV68复制的进一步鉴定发现，从感染细胞中输出的v-Cyclin零突变体和v-Cyclin CDK结合突变体存在严重缺陷。在这 R21的新应用，我们建议研究MHV68v-Cyclin在病毒从肺上皮细胞排出/释放中所起的作用。本研究的具体目的如下：1.研究v-Cyclin缺失突变体MHV68的出口表型：1.病毒粒子在wt和v-Cyclin突变体感染细胞中的定位；1.b病毒感染过程中v-Cyclin的细胞定位；1.c评估KSHV v-Cyclin缺失突变体在人脐静脉内皮细胞中的出口表型。目的2.分析无MHV68感染条件下v-Cyclin的功能：2.建立并鉴定可诱导表达v-Cyclin的细胞系；2.鉴定与v-Cyclin相互作用的伙伴。