Regulation of cholesterol biosynthesis by human parainfluenza virus type 1

1型人类副流感病毒对胆固醇生物合成的调节

• 批准号: 10307154
• 负责人: TORU TAKIMOTO
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-11-24 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307154
• 关键词: 3-hydroxy-3-methylglutaryl-coenzyme A; A549; Acute respiratory infection; Affect; Binding; Binding Proteins; Biogenesis; Bronchiolitis; Cause of Death; Cell Line; Cell Nucleus; Cell membrane; Cells; Child; Cholesterol; Cholesterol Homeostasis; Cholesterol Synthesis Inhibition; Coenzyme A; Cohort Analysis; Complex; Croup; Cytopathology; Data; Development; Endoplasmic Reticulum; Environment; Enzymes; Epidemiology; Epithelial Cells; Gene Expression; Genes; Genetic Transcription; Golgi Apparatus; Human; Immunoprecipitation; Impairment; Infant; Infection; Knock-out; Lead; Ligase; Mass Spectrum Analysis; Membrane Microdomains; Nucleocapsid; Oxidoreductase; Para-Influenza Virus Type 1; Para-Influenza Virus Type 3; Pathway interactions; Pattern; Phenotype; Play; Pneumonia; Population; Production; Proteins; Proteolysis; Public Health; RNA Viruses; Reaction; Regulation; Regulatory Element; Reporting; Research Proposals; Role; SCAP protein; Seasons; Sterol Biosynthesis Pathway; Sterols; Structural Protein; Time; Tracheal Epithelium; Transcriptional Regulation; Ubiquitination; Viral; Viral Genome; Viral Proteins; Viral Structural Proteins; Virion; Virus; Virus Assembly; airway epithelium; base; cholesterol biosynthesis; chronic infection; differential expression; innovation; insight; protein complex; recruit; respiratory; success; tool; trafficking; transcription factor; transcriptomics; ubiquitin ligase; ubiquitin-protein ligase

PROJECT SUMMARY/ABSTRACT Human parainfluenza virus type 1 (hPIV1) is one of the major causes of croup among young children and has a significant impact on public health. Humans are the only known host for hPIV1 infection, and the epidemiological patterns show seasonal and distinct biennial peaks of infection. It is not known how hPIV1 is maintained among the human population during the off season. So far, limited studies have been done regarding persistent hPIV1 infection in human airway cells. In this study, we investigated viral cytopathology, replication, and progeny virion release from human airway cells infected with hPIV1 to determine whether hPIV1 can establish persistent infection. We found that, unlike other respiratory RNA viruses, hPIV1 infection did not cause any cytopathic effects in human respiratory cells. Infected cells continue to produce viral structural proteins even at 15 days post infection. However, assembly and release of infectious virus decreased over time reaching 100-fold reduction on day 5 and remained low even at 15 dpi, showing that hPIV1 readily establishes quiescent infection. Interestingly, accumulation of large aggregates of viral nucleocapsid (RNP) was detected at late times post infection, suggesting impaired viral RNP trafficking and assembly of infectious virions at the plasma membrane. This phenotype is similar to what we observed in cholesterol-depleted cells, which specifically limited virus assembly and release. Strikingly, we found that hPIV1 infection reduces cholesterol levels by inhibiting expression of genes involved in sterol biosynthesis and ubiquitination and degradation of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), a rate-limiting enzyme in cholesterol biosynthesis. These data provide evidence for the first time that hPIV1 regulates cholesterol homeostasis, which likely contributes to the established quiescent infection in human respiratory epithelial cells. In this research proposal, we will elucidate the mechanism by which hPIV1 regulates cholesterol biosynthesis to establish a quiescent infection. Especially, we will analyze the role of Insig-1/2 proteins, which are known to be a key regulator for both transcriptional regulation and ubiquitination of HMGCR. Our hypothesis is that hPIV1 manipulates cellular cholesterol levels by targeting key regulators involved in cholesterol biosynthesis. We will identify the cellular and viral proteins responsible for hPIV1-induced inhibition of cholesterol synthesis, and analyze their molecular interactions. We will also determine how it contributes to quiescent infection of hPIV1. Success of this proposal will provide key data regarding how hPIV1 controls the cellular environment to establish quiescent infection.

项目摘要/摘要 人类1型副流感病毒(HPIV1)是引起幼儿哮喘的主要原因之一，具有 对公众健康产生重大影响。人类是唯一已知的hPIV1感染宿主，流行病学 模式显示了季节性和明显的两年一次的感染高峰。目前尚不清楚hPIV1是如何在 淡季的人口数量。到目前为止，关于持续性hPIV1的研究有限 人类呼吸道细胞的感染。在这项研究中，我们研究了病毒的细胞病理学、复制和子代病毒粒子。 从感染hPIV1的人呼吸道细胞中释放，以确定hPIV1是否可以建立持续感染。 我们发现，与其他呼吸道rna病毒不同，hPIV1感染不会对人类造成任何细胞病变。 呼吸系统细胞。即使在感染后15天，受感染的细胞仍会继续产生病毒结构蛋白。然而， 传染性病毒的组装和释放随着时间的推移而减少，在第5天减少了100倍，并保持在 即使在15dpi时也很低，表明hPIV1很容易建立静止性感染。有趣的是，大量的 病毒核衣壳聚合体(RNP)在感染后的后期被检测到，表明病毒RNP受损 侵染性病毒粒子在质膜上的运输和组装。这种表型与我们的 在胆固醇耗尽的细胞中观察到，这特别限制了病毒的组装和释放。令人惊讶的是，我们发现 HPIV1感染通过抑制参与类固醇生物合成的基因的表达来降低胆固醇水平 3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)的泛素化和降解 胆固醇的生物合成。这些数据首次提供了hPIV1调节胆固醇的证据 动态平衡，这可能是人类呼吸道上皮细胞已建立的静止感染的原因。在……里面 本研究建议，我们将阐明hPIV1调节胆固醇生物合成的机制 建立一种静止性感染。特别是，我们将分析Insig-1/2蛋白的作用，它是一种已知的 HMGCR转录调控和泛素化的关键调控因子。我们的假设是hPIV1 通过靶向参与胆固醇生物合成的关键调节因子来操纵细胞胆固醇水平。我们会 鉴定与hPIV1抑制胆固醇合成有关的细胞和病毒蛋白，并分析 它们的分子相互作用。我们还将确定它如何促进hPIV1的静止性感染。取得的成功 该提案将提供有关hPIV1如何控制蜂窝环境以建立静止状态的关键数据 感染。