Dynamic virus-driven remodeling of ER-mitochondria contacts

内质网-线粒体接触的动态病毒驱动重塑

• 批准号: 10608035
• 负责人: Ileana M. Cristea
• 金额: $ 41.94万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10608035
• 关键词: Acetylation; Activities of Daily Living; Address; Area; Binding; Bioenergetics; Biogenesis; Biological Assay; Biological Process; Cell Respiration; Cell Survival; Cells; Coupled; Crista ampullaris; Cryo-electron tomography; Cues; Cytomegalovirus; Cytomegalovirus Infections; Daughter; Depressed mood; Electron Microscopy; Electron Transport; Event; Herpesviridae; Hour; Human; Hybrids; Image; Immune response; In Situ; Infection; Influenza; Inner mitochondrial membrane; Lamins; Ligation; Link; Lipids; Lysosomes; Mass Spectrum Analysis; Membrane; Metabolic; Metabolic Diseases; Metabolism; Microscopy; Mitochondria; Modeling; Molecular; Molecular Virology; Morphogenesis; Nuclear; Organelles; Outcome; Output; Oxidative Phosphorylation; Oxygen Consumption; Pathology; Pathway interactions; Peripheral; Population; Production; Proteins; Proteomics; Regulation; Reporting; Research; Resolution; Respiration; Role; Shapes; Signal Transduction; Site; Structure; Structure-Activity Relationship; Viral; Viral Pathogenesis; Viral Physiology; Viral Proteins; Virion; Virus; Virus Diseases; Work; betacoronavirus; genetic manipulation; interdisciplinary approach; light microscopy; lipidomics; live cell microscopy; metabolomics; mitochondrial membrane; peroxisome; recruit; release of sequestered calcium ion into cytoplasm; respiratory; spatiotemporal; virology

Viruses have evolved elegant strategies to manipulate host cell machinery and rewire core cellular pathways to facilitate productive infection, including enhancing metabolic output and maintaining cell viability. To accomplish this, viruses exert an extensive network of dynamic molecular interactions with cellular organelles. As the functions of organelles are intimately associated with the regulation of their composition, shape, and localization, the control of organelle structure-function relationships is at the core of clarifying the outcome of an infection. While many examples of virus-induced organelle remodeling have been described, very little is understood about how organelle structures engender specific functions. Our lab has characterized a previously unrecognized aspect of viral infection, which is that human viruses globally control organelle remodeling by dramatically rewiring inter- and intra-organelle membrane contact sites (MCS). Using a hybrid quantitative proteomics and super resolution microscopy approach, we demonstrated exquisite reorganization in MCS networks engaged by a broad range of human viruses, including both ancient (herpesviruses) and rapidly adapting (influenza and beta- coronavirus) viruses. We further discovered that infection with the ubiquitous herpesvirus human cytomegalovirus (HCMV) triggers a new specialized MCS structure, mitochondria-ER encapsulations that we termed MENC. We determined that HCMV infection drives predominantly fission at the mitochondrial periphery, and that the fragmented mitochondria enter MENCs and retain their bioenergetic activity. How the infection induces MENC formation and the function of this newly reported structure remain unknown. We propose that MENCs provide a unifying explanation for the longstanding paradox of how certain viruses such as HCMV increase mitochondrial bioenergetic output, despite inducing mitochondrial fragmentation. Our central hypothesis is that HCMV remodels inter- and intra-organelle connections, generating MENCs, which act to protect and stabilize the bioenergetic capacity of fragmented mitochondria. Using a multidisciplinary approach that combines molecular virology with cutting-edge approaches in quantitative proteomics, live super resolution microscopy, ultrastructural electron microscopy, metabolomics, and lipidomics, in Aim 1, we will define the mechanisms underlying the formation and function of MENCs during HCMV infection. In Aim 2, we will establish what signaling cues from HCMV-induced three-way contacts among the ER, mitochondria, and lysosome stimulate peripheral mitochondria fission and elevate bioenergetic respiration. In Aim 3, we will characterize the viral factors that coordinate ER-mitochondria MCS rewiring. Collectively, our study will link newly discovered aspects of virus-orchestrated MCS networking to new two-way and three-way organelle structure-function relationships that underlie fundamental cellular mechanisms, including mitochondrial bioenergetics and autophagic turnover. In doing so, our study will open research areas in how viruses exploit the functional capacities of remodeled organelles for infection, which have broad implications for viral pathogenesis and metabolic disorders.

病毒已经进化出优雅的策略来操纵宿主细胞机器并重新连接核心细胞通路 促进生产性感染，包括增强代谢输出和维持细胞活力。为了完成 因此，病毒与细胞器发挥了广泛的动态分子相互作用网络。作为 细胞器的功能与其组成、形状和定位的调节密切相关， 细胞器结构-功能关系的控制是阐明感染结果的核心。 虽然已经描述了许多病毒诱导的细胞器重塑的例子，但人们对它知之甚少。 细胞器结构如何产生特定功能。我们的实验室发现了一种以前未被识别的特征 病毒感染的一个方面，即人类病毒通过显着地控制细胞器重塑 重新布线细胞器间和细胞器内膜接触位点（MCS）。使用混合定量蛋白质组学和 超分辨率显微镜方法，我们展示了 MCS 网络的精致重组 多种人类病毒，包括古老的病毒（疱疹病毒）和快速适应的病毒（流感病毒和β-病毒） 冠状病毒）病毒。我们进一步发现，感染人类普遍存在的疱疹病毒 巨细胞病毒 (HCMV) 触发一种新的专门 MCS 结构，即线粒体-ER 封装，我们将其 称为 MENC。我们确定 HCMV 感染主要驱动线粒体外周的裂变， 碎片化的线粒体进入 MENC 并保留其生物能活性。怎么传染的 诱导 MENC 形成，但这种新报道的结构的功能仍然未知。我们建议 MENC 为某些病毒（如 HCMV）如何长期存在的悖论提供了统一的解释。 尽管诱导线粒体断裂，但仍增加线粒体生物能输出。我们的中心假设 HCMV 重塑细胞器间和细胞内连接，产生 MENC，其作用是保护和 稳定碎片线粒体的生物能能力。使用多学科方法结合 分子病毒学与定量蛋白质组学、实时超分辨率显微镜、 超微结构电子显微镜、代谢组学和脂质组学，在目标 1 中，我们将定义其机制 HCMV 感染期间 MENC 的形成和功能的基础。在目标 2 中，我们将确定什么 HCMV 诱导的 ER、线粒体和溶酶体之间三向接触的信号线索刺激 外周线粒体裂变并增强生物能呼吸。在目标 3 中，我们将描述病毒的特征 协调 ER 线粒体 MCS 重连的因素。总的来说，我们的研究将把新发现的方面联系起来 病毒编排的MCS网络到新的双向和三向细胞器结构-功能关系 这是基本细胞机制的基础，包括线粒体生物能学和自噬周转。 在此过程中，我们的研究将开辟病毒如何利用重塑的功能能力的研究领域。 感染的细胞器，对病毒发病机制和代谢紊乱具有广泛的影响。