Membrane and lipidome dynamics during enterovirus infection

肠道病毒感染期间的膜和脂质组动力学

基本信息

批准号：
10751143
负责人：
David Aponte-Diaz
金额：
$ 3.57万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10751143
关键词：
Ablation Achievement Anabolism Antibodies Antiviral Agents Biochemical Biogenesis Biological Process Categories Cell Line Cell Nucleus Cell membrane Cell physiology Cells Cellular Membrane Child Cytoplasm Data Defect Diglycerides Disease Enterovirus Enterovirus Infections Enzymes Event FOS Protein FOS gene Family Fatty Acids Genes Genetic Determinism Genetic Recombination Genetic Screening Genetic Transcription Genome Goals Hela Cells Human poliovirus Immunofluorescence Immunologic Impairment Infection Innate Immune Response Integration Host Factors Investigation Knock-out Lecithin Life Cycle Stages Lipids Luciferases Mammalian Cell Mass Spectrum Analysis Mediating Membrane Modeling Modification Morbidity - disease rate Mutation Neurologic Null Lymphocytes Organelles Pathway interactions Pattern recognition receptor Phenotype Phosphatidate Phosphatase Phosphatidic Acid Phosphatidylcholine Biosynthesis Phospholipids Production Protein Dephosphorylation Protein Isoforms Proteins Public Health Publishing RNA RNA Viruses Reporter Research Risk Role Symptoms Transfection Translations Vaccines Variant Very Long Chain Fatty Acid Vesicle Viral Viral Genome Viral Physiology Viral Proteins Virion Virulent Virus Virus Assembly Virus Diseases Virus Replication Work biophysical properties falls inhibitor insight lipid biosynthesis lipid metabolism lipidome lipidomics membrane biogenesis metaplastic cell transformation new outbreak non-genomic novel posttranscriptional prevent programs respiratory spatiotemporal stem cells trafficking transcription factor

项目摘要

Summary/Abstract Enterovirus infections are a significant public health burden worldwide. Nearly 15 million enterovirus infections occur each year. While infection predominantly elicits mild-symptom disease, some enterovirus strains may confer severe respiratory and neurological illnesses with higher morbidity in children. There are no current antiviral therapeutics to thwart enteroviral infections. All positive-strand RNA viruses, even non-enveloped poliovirus (PV)-like enteroviruses, require host membranes for multiplication. These viruses remodel the host lipidome to create virus-induced membranes with unique phospholipid composition, thereby conferring unique biochemical and biophysical properties upon these membranes that enable distinct biological functions. PV- related enteroviruses and likely many other viruses fall into this category. Remarkably, during PV infection, translation of the infecting RNA is sufficient to induce cellular transformations before genome replication or host transcription is engaged. This observation suggests that post-transcriptional and/or post-translational mechanisms exist in the mammalian cell cytoplasm to reprogram phospholipid biosynthesis and membrane biogenesis minutes after infection and that “hubs,” which can be co-opted by PV, control these mechanisms. Our research aims to illuminate mechanisms regulating membrane biogenesis, function, and trafficking in cells by understanding how PV co-opts these mechanisms. PV 3CD induces multiple phospholipids during infection. To investigate the mechanism, we expressed a tagged 3CD in HeLa cells and exploited the tag to isolate 3CD- associated proteins. We identified 3CD-associated proteins using mass spectrometry. We combined our data with genetic screens and published data on known interactions to arrive at a hypothetical model for lipid induction. By evaluating proteins with two or three degrees of separation from interactions discovered empirically, c-Fos appeared. Studies have demonstrated a direct role of c-Fos in activating enzymes that perform rate-limiting steps in phospholipid biosynthesis and membrane biogenesis. We hypothesize that PV modulates c-Fos to form PV-induced membranes of unique phospholipid composition by exploiting multiple lipid biosynthetic pathways. Thus Aim 1 will evaluate PV-mediated c-Fos modulation for the induction of phospholipid biosynthesis and membrane biogenesis. Furthermore, our preliminary data evaluating the lipidome of PV-infected cells shows the induction of very long-chain fatty acids synthesis. Only one isoform in the Elongation of Very Long-Chain Fatty Acids family of enzymes (ELOVL4) produces these fatty acids. We hypothesize that PV viral factors hijack cellular pathways (namely ELOVL4) to produce long fatty acids that stabilize virus-containing vesicles during infection. Thus Aim 2 will characterize the dynamics, mechanisms, and functions of host-lipidome remodeling during PV infection. These studies will not only elucidate pathways governing membrane dynamics during viral infection but also provide insight into how these pathways are regulated during normal cellular physiology—presenting attractive target candidates for developing antivirals.

摘要/摘要肠道病毒感染是全球近 1500 万肠道病毒感染的重大公共卫生负担。虽然感染主要引起轻微症状的疾病，但某些肠道病毒株可能每年都会发生。会导致严重的呼吸系统和神经系统疾病，儿童发病率更高。阻止肠道病毒感染的抗病毒疗法。所有正链 RNA 病毒，甚至是无包膜病毒。脊髓灰质炎病毒 (PV) 样肠道病毒需要宿主膜才能繁殖，这些病毒会重塑宿主。脂质组创建具有独特磷脂组成的病毒诱导膜，从而赋予独特的这些膜具有独特的生物化学和生物物理特性，具有独特的生物功能。相关的肠道病毒和可能的许多其他病毒都属于这一类。值得注意的是，在真性红斑狼疮感染期间，感染RNA的翻译足以在基因组复制之前诱导细胞转化或这一观察表明转录后和/或翻译后参与。哺乳动物细胞质中存在重新编程磷脂生物合成和细胞膜的机制感染后几分钟的生物发生以及可以被PV选择的“中枢”控制这些机制。我们的研究旨在阐明调节细胞膜生物发生、功能和运输的机制通过了解 PV 如何利用这些机制，PV 3CD 在感染过程中诱导多种磷脂。为了研究其机制，我们在 HeLa 细胞中表达了带标签的 3CD，并利用该标签分离了 3CD- 我们使用质谱法鉴定了 3CD 相关蛋白。通过基因筛选和已知相互作用的已发表数据，得出脂质的假设模型通过评估与发现的相互作用具有两级或三级分离的蛋白质。根据经验，c-Fos 的出现已证明 c-Fos 在激活酶方面具有直接作用。在磷脂生物合成和膜生物发生中执行限速步骤我们突袭了PV。通过利用多种机制，调节 c-Fos 形成具有独特磷脂成分的 PV 诱导膜因此，目标 1 将评估 PV 介导的 c-Fos 调节以诱导此外，我们的初步数据评估了磷脂生物合成和膜生物发生。 PV 感染细胞的脂质组显示仅诱导一种长链脂肪酸合成。长链脂肪酸延长酶家族 (ELOVL4) 产生这些脂肪酸。认为 PV 病毒因子劫持细胞通路（即 ELOVL4）以产生长脂肪酸，因此，目标 2 将表征感染过程中含有病毒的囊泡的动力学、稳定机制，这些研究不仅会阐明PV感染期间宿主脂质体重塑的功能。控制病毒感染期间的膜动力学，同时也提供了对这些途径如何发挥作用的深入了解在正常细胞生理过程中受到调节——为开发抗病毒药物提供了有吸引力的候选靶点。