SUMO and ubiquitin modifications in henipavirus matrix trafficking and function

亨尼帕病毒基质运输和功能中的相扑和泛素修饰

• 批准号: 9159123
• 负责人: Benhur Lee
• 金额: $ 43.73万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-24 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9159123
• 关键词: Affect; Animals; Automobile Driving; Behavior; Biology; Cell Nucleus; Cell membrane; Cells; Cellular biology; China; Chiroptera; Complex; Cytoplasm; Data; Data Set; Disease; Disease Outbreaks; Dominant-Negative Mutation; Ectopic Expression; Family; Fruit; Goals; Hendra Virus; Henipavirus; Human; Infection; Life; Life Cycle Stages; Ligase; Link; Livestock; Lysine; Mass Spectrum Analysis; Mediating; Membrane; Modification; Nuclear; Nuclear Matrix-Associated Proteins; Paramyxovirus; Pathway interactions; Peptide Hydrolases; Phenocopy; Phenotype; Plasma; Play; Pneumonia; Population; Post-Translational Protein Processing; Proteasome Inhibition; Proteins; Proteomics; Recombinants; Risk; Role; Site; System; Therapeutic; Therapeutic Intervention; Transfection; Ubiquitin; Ubiquitination; Viral; Virion; Virulent; Virus; Virus Assembly; Virus Diseases; Virus Replication; Work; fitness; high risk; mortality; multicatalytic endopeptidase complex; multiple myeloma M Protein; mutant; novel; overexpression; pathogen; protein transport; reverse genetics; targeted treatment; trafficking; transmission process; ubiquitin-protein ligase

The Paramyxovirus family contains numerous human and animal pathogens of global concern. Nipah (NiV) and Hendra (HeV) viruses of the Henipavirus (HNV) genus are the most virulent paramyxoviruses that infect humans. Zoonotic transmission of NiV and HeV from their natural fruit bat reservoirs to humans can result in mortality rates in excess of 90%. The recent discovery of novel clades of HNV across the globe highlights the possible global emergence of these zoonotic viruses. A better understanding of the basic pathobiology of highly pathogenic viruses is critical for developing therapeutic options. The Paramyxovirus matrix (M) protein is critical to viral assembly and budding. Although paramyxoviruses replicate entirely in the cytoplasm, we found that early during infection, NiV-M protein is first targeted to the nucleus prior to ubiquitination and subsequent localization to the plasma membrane. This ubiquitin-regulated nuclear-cytoplasmic trafficking of M is critical for its ability to mediate viral assembly and budding, and is conserved across viruses from several Paramyxovirus genera. This application is spurred by a confluence of further preliminary data suggesting that NiV-M is SUMOylated, and that the SUMOylation state of the cell affects NiV-M's function. Our overall goal is to determine the role of SUMO and ubiquitin modifications in matrix trafficking and function. Our objective is to understand how cognate SUMO and ubiquitin proteasome pathway components coordinately regulate the complex intracellular trafficking behavior of HNV-M proteins, and to uncover the functional role(s) that these post-translational modifications play in the virus replicative life cycle. Our driving hypothesis is that specific SUMO and ubiquitin proteasome pathway components coordinate the spatial-temporal control of the matrix protein's nuclear sojourn, which enables the proper membrane targeting of matrix that is critical for virus assembly and budding. Understanding the basic cell biology of HNV-M might reveal host-pathogen interactions that could be targeted for therapeutic interventions. To interrogate our driving hypothesis, we propose the following Specific Aims: AIM 1: Determine which lysines in HNV-M are modified. AIM 2: Interrogate the consequences of HNV-M interactions with E3 Ub-ligases. AIM 3: Evaluate the role of SUMOylation and Nup358/RanBP2 (a SUMO E3 ligase) in the nuclear trafficking and function of HNV-M proteins. AIM 4: Define the functional roles for matrix post-translational modifications in relation to viral replication and pathogenic fitness.

副粘病毒家族包含许多全球关注的人类和动物病原体。亨尼帕病毒 (HNV) 属的尼帕 (NiV) 和亨德拉 (HeV) 病毒是感染人类的​​毒性最强的副粘病毒。 NiV 和 HeV 从天然果蝠宿主传播给人类的人畜共患病可导致超过 90% 的死亡率。最近在全球范围内发现的新的 HNV 分支凸显了这些人畜共患病毒可能在全球范围内出现。更好地了解高致病性病毒的基本病理学对于开发治疗方案至关重要。副粘病毒基质 (M) 蛋白对于病毒组装和出芽至关重要。尽管副粘病毒完全在细胞质中复制，但我们发现在感染早期，NiV-M 蛋白首先靶向细胞核，然后泛素化并随后定位到质膜。 M 的这种泛素调节的核细胞质运输对于其介导病毒组装和出芽的能力至关重要，并且在多个副粘病毒属的病毒中是保守的。这一应用受到进一步初步数据的融合的推动，这些数据表明 NiV-M 被 SUMO 化，并且细胞的 SUMO 化状态影响 NiV-M 的功能。我们的总体目标是确定 SUMO 和泛素修饰在基质运输和功能中的作用。我们的目标是了解同源 SUMO 和泛素蛋白酶体途径成分如何协调调节 HNV-M 蛋白复杂的细胞内运输行为，并揭示这些翻译后修饰在病毒复制生命周期中发挥的功能作用。我们的驱动假设是，特定的 SUMO 和泛素蛋白酶体途径成分协调基质蛋白核停留的时空控制，从而实现基质的正确膜靶向，这对于病毒组装和出芽至关重要。了解 HNV-M 的基本细胞生物学可能会揭示宿主与病原体之间的相互作用，从而可以作为治疗干预的目标。为了质疑我们的驱动假设，我们提出以下具体目标： 目标 1：确定 HNV-M 中的哪些赖氨酸被修饰。目标 2：探讨 HNV-M 与 E3 Ub 连接酶相互作用的后果。目标 3：评估 SUMO 化和 Nup358/RanBP2（一种 SUMO E3 连接酶）在 HNV-M 蛋白的核运输和功能中的作用。目标 4：定义与病毒复制和致病适应性相关的基质翻译后修饰的功能作用。