CAREER: Murine Gammaherpesvirus 68 Persistence and Interaction With Host Chemokine Networks

职业奖：小鼠γ疱疹病毒68持久性和与宿主趋化因子网络的相互作用。病毒与宿主的进化相互作用反映了不同的适应策略。疱疹病毒是一种共同进化策略的例证，在这种策略中，宿主的先天和适应性免疫防御以及细胞信号通过病毒诱导的调节来平衡，这种调节通常会导致体内平衡的改变，从而促进病毒与多细胞宿主的永久结合。在保持这种平衡的同时，主人的健身成本通常很少。疱疹病毒的大DNA基因组编码了许多辅助基因，这些基因对病毒复制是不必要的，但通过调节宿主反应，对病毒的持久性至关重要。γ疱疹病毒能够在淋巴样细胞中持续存在，它包含了一系列令人印象深刻的基因，这些基因显然是通过分子窃取宿主基因获得的，这些基因与免疫识别和应答、细胞周期控制、补体介导的杀伤和凋亡应答有关。在γ -2疱疹病毒（或rhadino病毒）的遗传库中，突出的是编码与宿主趋化因子网络相互作用产物的基因，这些趋化因子网络在稳态和炎症条件下的细胞交通调节中都是至关重要的。在一些病毒基因组中保守的是一个g蛋白偶联受体（GPCR）基因，该基因编码一个与CXCR2最密切相关的cxc趋化因子受体同源物。这些研究将在大量学生参与的情况下进行伽马疱疹病毒潜伏期和持久性的研究，以及对小鼠γ疱疹病毒68 （MHV-68）的ORF74编码的病毒CXC趋化因子受体的功能分析。利用强大的遗传工具，MHV-68与小鼠宿主的相互作用可以很容易地进行研究，使该系统非常适合于推进对γ疱疹病毒生物学的理解，包括γ疱疹病毒潜伏期和持久性的机制。MHV-68基因组已经完全测序，可以使用常规的体外重组方法进行操作。遗传定义的小鼠菌株（包括转基因菌株和具有特定基因敲除的菌株）的可用性为使用病毒和宿主遗传学操作来查询病毒-宿主关联的性质提供了无与伦比的机会。这些关于MHV-68持久性和与宿主趋化因子网络相互作用的研究将利用这种病毒-宿主系统的优势。已知的伽马疱疹病毒-宿主相互作用的有趣性质，适当的实验方法的优雅，包括分子生物学标准的实验方法，以及大量未解决的问题，共同提供了一个研究框架，非常适合整合到涉及研究生，本科生和大学预科学生及其教师的教育活动中。使用普通分子生物学方法克隆MHV-68基因的研究将继续为入门和高级分子生物学实验室课程提供探究式的背景。利用首席研究员在构建重组rhadino病毒以研究病毒背景下基因功能方面的丰富经验，研究生项目将包括构建有助于理解病毒-宿主相互作用的MHV-68重组病毒。研究生和本科生将使用克隆病毒基因表达构建体在体外进行MHV-68基因功能的研究。与地区高中学生及其老师的持续互动将为选定的学生提供机会，让他们参与小型的、定义明确的实验室研究项目，利用基本的分子生物学方法。

CAREER Award: Murine gammaherpesvirus 68 persistence and interaction with hostchemokine networks.The evolutionary interactions of viruses and their hosts reflect diverse adaptive strategies.Herpesviruses exemplify a co-evolutionary strategy in which host innate and adaptive immune defenses and cellular signalling are balanced by virus induced modulations that typically result in an altered homeostasis facilitating permanent association of the virus and the multicellular host. While this balance is maintained, there is usually little cost in fitness for the host. The large DNA genomes of herpesviruses encode numerous accessory genes unnecessary for viral replication but critically involved in virus persistence through modulation of host responses. Gammaherpesviruses, which are capable of persistence in lymphoid cells, contain a particularly impressive variety of genes apparently acquired through molecular piracy of host genes relevant to immune recognition and response, cell cycle control, complement-mediated killing, and apoptotic responses. Prominent within the genetic repertoire of gamma-2-herpesviruses (or rhadinoviruses) are genes encoding products that interact with host chemokine networks which are critical in regulation of cell traffic under both homeostatic and inflammatory conditions. Conserved within several rhadinovirus genomes is a G-protein-coupled receptor (GPCR) gene encoding a putative CXC-chemokine receptor homolog most closely related to CXCR2. Studies of gammaherpesvirus latency and persistence together with functional analysis of the viral CXC chemokine receptor encoded by ORF74 of murine gammaherpesvirus 68 (MHV-68) will be pursued in these studies with extensive student participation. Interaction of MHV-68 with murine hosts is highly accessible to investigation usingpowerful genetic tools making this system ideally suited for advancing understanding ofgammaherpesvirus biology including mechanisms of gammaherpesvirus latency and persistence.The MHV-68 genome has been fully sequenced and can be manipulated using conventional in vitro recombination methods. Availability of genetically defined mouse strains (including transgenic strains and those with specific gene knockouts) affords unparalleled opportunities to query the nature of the virus-host association using manipulations of both virus and host genetics. These investigations of MHV-68 persistence and interaction with host chemokine networks will exploit the advantages of this virus-host system. The intriguing nature of known gammaherpesvirus-host interactions, the elegance ofappropriate experimental methods including those that are standard to molecular biology, and the abundance of unanswered questions together provide a research framework that is well suited for integration into educational activities involving graduate students, undergraduates, and pre-college students and their teachers. Studies of cloned MHV-68 genes using common molecular biological methods will continue to provide an inquiry-based context for introductory and advanced molecular biology laboratory courses. Drawing upon the extensive experience of the principal investigator in construction of recombinant rhadinoviruses for study of gene function within the viral context, graduate student projects will include construction of MHV-68 recombinants useful for understanding virus-host interactions. Both graduate and undergraduate students will perform studies of MHV-68 gene function in vitro using cloned viral gene expression constructs. Ongoing interactions with regional high school students and their teacherswill provide opportunities for selected students to participate in small, well defined laboratory research projects utilizing basic molecular biological methods.