Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, US.
Department of Microbiology, University of Alabama Birmingham, Birmingham, AL, 35294, US.
Nat Commun. 2022 Aug 11;13(1):4720. doi: 10.1038/s41467-022-32488-6.
Membrane contact sites (MCSs) link organelles to coordinate cellular functions across space and time. Although viruses remodel organelles for their replication cycles, MCSs remain largely unexplored during infections. Here, we design a targeted proteomics platform for measuring MCS proteins at all organelles simultaneously and define functional virus-driven MCS alterations by the ancient beta-herpesvirus human cytomegalovirus (HCMV). Integration with super-resolution microscopy and comparisons to herpes simplex virus (HSV-1), Influenza A, and beta-coronavirus HCoV-OC43 infections reveals time-sensitive contact regulation that allows switching anti- to pro-viral organelle functions. We uncover a stabilized mitochondria-ER encapsulation structure (MENC). As HCMV infection progresses, MENCs become the predominant mitochondria-ER contact phenotype and sequentially recruit the tethering partners VAP-B and PTPIP51, supporting virus production. However, premature ER-mitochondria tethering activates STING and interferon response, priming cells against infection. At peroxisomes, ACBD5-mediated ER contacts balance peroxisome proliferation versus membrane expansion, with ACBD5 impacting the titers of each virus tested.
膜接触位点 (MCSs) 将细胞器连接起来，以协调跨时空的细胞功能。尽管病毒在复制周期中重塑细胞器，但在感染过程中，MCSs 仍然在很大程度上未被探索。在这里，我们设计了一种靶向蛋白质组学平台，用于同时测量所有细胞器的 MCS 蛋白，并通过古老的β疱疹病毒人类巨细胞病毒 (HCMV) 定义功能性病毒驱动的 MCS 改变。与超分辨率显微镜的整合以及与单纯疱疹病毒 (HSV-1)、甲型流感病毒和β冠状病毒 HCoV-OC43 感染的比较表明，接触调节具有时间敏感性，允许从抗病毒细胞器功能切换到促病毒细胞器功能。我们发现了一种稳定的线粒体-内质网包裹结构 (MENC)。随着 HCMV 感染的进展，MENCs 成为主要的线粒体-内质网接触表型，并依次招募连接伙伴 VAP-B 和 PTPIP51，支持病毒产生。然而，内质网-线粒体过早连接会激活 STING 和干扰素反应，使细胞对感染产生初步反应。在过氧化物酶体中，ACBD5 介导的内质网接触平衡过氧化物酶体增殖与膜扩张，ACBD5 影响每种测试病毒的滴度。
