Defining the regulation of mitochondrial bioenergetics during virus infection

项目总结/摘要。 人巨细胞病毒（HCMV）是一种疱疹病毒，感染世界上超过50%的人口， 造成个体终身感染。巨细胞病毒感染是一个主要的问题，在个人受损或 幼稚的免疫系统，因为它可以导致一系列疾病，包括耳聋，呼吸系统疾病和器官 失败此外，HCMV由于其在免疫系统中的作用，近年来引起了越来越多的关注。 慢性疾病的出现和发展，如心血管疾病和癌症。一个显著特点 巨细胞病毒感染的主要原因是细胞代谢的全球重新布线， 前体和复制的能量。HCMV感染时细胞代谢的失调， 它的复制所必需的，并已被链接到它的许多病理，包括它的肿瘤调节 容量然而，我们目前缺乏对代谢改变的机制的理解 在感染过程中观察到。我们最近发现，线粒体酶sirtuin 4（SIRT 4）是一种有效的 抗病毒因子在HCMV感染中的作用此外，我们建立了SIRT 4作为第一个已知的哺乳动物细胞， 脂酰胺酶，从丙酮酸脱氢酶中去除必需的翻译后修饰脂酰化 复杂.这一发现指出SIRT 4是细胞代谢的关键调节因子，但SIRT 4如何发挥其作用， 抗病毒功能仍然未知。我推测SIRT 4在HCMV感染过程中的宿主防御中起作用 通过对抗病毒引起的细胞代谢变化。进一步支持SIRT 4在抗病毒治疗中的关键作用 当HCMV感染时，我发现HCMV已经获得了一种抑制其功能的机制。我的初步结果 证明SIRT 4被先前未表征的病毒蛋白pUL 13靶向抑制。在我 在这个建议中，我将讨论病毒与宿主相互作用的两个方面。在目标1中，分子病毒学， 显微镜、蛋白质组学和代谢组学将被用于定义SIRT 4介导的防御机制 抗HCMV感染。我将确定哪些特定的SIRT 4酶活性是抗病毒所必需的， 反应在目标2中，我将揭示pUL 13如何抑制SIRT 4，以及表征其在调节SIRT 4中的功能。 细胞代谢和线粒体生物能量学。作为一项长期目标，阐明 SIRT 4和pUL 13之间相互作用可以帮助解释HCMV如何诱导代谢变化， 疾病这些知识可以指向恢复代谢健康和治疗HCMV的治疗靶点。 相关病理这项研究将在普林斯顿大学分子生物学系进行， 该计划以其多学科研究和研究生培训的支持环境而闻名， 职业发展。

PROJECT SUMMARY/ABSTRACT. Human cytomegalovirus (HCMV) is a ß-herpesvirus that infects over 50% of the world’s population and establishes lifelong infection in individuals. HCMV infection is a major concern in individuals with impaired or naïve immune systems, as it can lead to a range of diseases, including deafness, respiratory disease, and organ failure. Additionally, HCMV has garnered increased interest in recent years due to its implication in the emergence and progression of chronic diseases, such as cardiovascular disease and cancer. A striking feature of HCMV infection is the global rewiring of cellular metabolism for the increased production of biosynthetic precursors and energy for replication. The dysregulation of cellular metabolism during HCMV infection is necessary for its replication and has been linked to many of its pathologies, including its oncomodulatory capacity. However, we currently lack an understanding of the mechanisms underlying the metabolic alterations observed during infection. We recently discovered that the mitochondrial enzyme sirtuin 4 (SIRT4) is a potent antiviral factor during HCMV infection. Furthermore, we established SIRT4 as the first known mammalian cellular lipoamidase, removing the essential posttranslational modification lipoylation from the pyruvate dehydrogenase complex. This discovery points to SIRT4 as a critical regulator of cellular metabolism, but how SIRT4 exerts its antiviral function remains unknown. I hypothesize that SIRT4 functions in host defense during HCMV infection by opposing viral-induced changes in cellular metabolism. Further supporting the critical role of SIRT4 in antiviral response, I discovered that HCMV has acquired a mechanism to suppress its functions. My preliminary results demonstrate that SIRT4 is targeted for inhibition by the previously uncharacterized viral protein, pUL13. In my proposal, I will address both sides of this virus-host interplay. In Aim 1, a combination of molecular virology, microscopy, proteomics and metabolomics will be used to define SIRT4-mediated mechanisms of defense against HCMV infection. I will determine which specific SIRT4 enzymatic activities are required for antiviral response. In Aim 2, I will uncover how pUL13 inhibits SIRT4, as well as characterize its function in regulating cellular metabolism and mitochondrial bioenergetics. As a long-term objective, elucidating the functional interaction between SIRT4 and pUL13 can help explain how HCMV induces metabolic changes that promote disease. This knowledge can point to therapeutic targets for restoring metabolic health and for treating HCMV- linked pathologies. This research will take place in the Molecular Biology Department of Princeton University, a program known for its multidisciplinary research and supportive environment for graduate student training and career development.