Defining the regulation of mitochondrial bioenergetics during virus infection

项目摘要/摘要。 人巨细胞病毒(HCMV)是一种感染世界50%以上人口的疱疹病毒， 在个人中建立终身感染。人巨细胞病毒感染是患有受损或 天真的免疫系统，因为它会导致一系列疾病，包括耳聋、呼吸系统疾病和器官 失败了。此外，近年来，人巨细胞病毒引起了越来越多的关注，因为它在 慢性疾病的出现和发展，如心血管疾病和癌症。一个引人注目的特征 巨细胞病毒感染是细胞代谢的全球重新布线，以增加生物合成的产生 复制的前体和能量。人巨细胞病毒感染过程中细胞代谢紊乱 对它的复制是必要的，并与它的许多病理联系在一起，包括它的肿瘤调节 容量。然而，我们目前对代谢改变的机制缺乏了解。 在感染期间观察到的。我们最近发现线粒体酶sirtuin 4(Sirtuin 4)是一种有效的 人巨细胞病毒感染过程中的抗病毒因子。此外，我们建立了SIRT4作为第一个已知的哺乳动物细胞 从丙酮酸脱氢酶中去除必要的翻译后修饰的硫代化 很复杂。这一发现表明SIRT4是细胞代谢的关键调节因子，但SIRT4是如何发挥其作用的 抗病毒功能尚不清楚。我推测SIRT4在人巨细胞病毒感染过程中起宿主防御作用 通过反对病毒引起的细胞新陈代谢变化。进一步支持SIRT4在抗病毒中的关键作用 反应后，我发现巨细胞病毒已经获得了一种抑制其功能的机制。我的初步结果 证明SIRT4是以前未确定特征的病毒蛋白pUL13的抑制靶点。在我的 提案中，我将讨论病毒与宿主相互作用的两个方面。在目标1中，结合了分子病毒学， 显微镜、蛋白质组学和代谢组学将用于确定SIRT4介导的防御机制 抗人巨细胞病毒感染。我将确定哪些特定的SIRT4酶活性是抗病毒所必需的 回应。在目标2中，我将揭示pUL13是如何抑制SIRT4的，并表征其调节功能 细胞代谢和线粒体生物能量学。作为一个长期目标，阐明功能 SIRT4和pUL13之间的相互作用有助于解释HCMV如何诱导代谢变化，从而促进 疾病。这种知识可以指向恢复代谢健康和治疗HCMV的治疗目标- 相互关联的病理。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养和支持环境而闻名 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.