Proteomic and molecular analysis of early events in HSV assembly

HSV 组装早期事件的蛋白质组学和分子分析

• 批准号: 8239988
• 负责人: DUNCAN W. WILSON
• 金额: $ 56.07万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8239988
• 关键词: ANXA2 gene; Address; Amino Acids; Binding; Binding Proteins; Biochemical; Biochemistry; Biological; Biology; Blindness; Capsid; Cell Nucleus; Cytoplasm; DNA Packaging; Data; Disease; Encephalitis; Event; Eye; Generations; Genital system; Herpes Simplex Infections; Herpesviridae; Herpesvirus 1; Lead; Lesion; Location; Mass Spectrum Analysis; Membrane; Methodology; Modeling; Molecular; Molecular Analysis; Neuraxis; Nuclear; Nuclear Inner Membrane; Nuclear Outer Membrane; Oral; Pharmaceutical Preparations; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; Protocols documentation; Reporter; Role; Simplexvirus; Site; Sorting - Cell Movement; Structure; Testing; Therapeutic; United States; Viral; Viral Proteins; Virion; Virus; Virus Assembly; Virus Diseases; Virus Replication; env Gene Products; particle; polypeptide; protein complex; public health relevance; research study; trafficking

DESCRIPTION (provided by applicant): All Herpes viruses assemble their DNA-packaged capsids in the infected cell nucleus. These capsids then undergo a process unique in biology: they bud into the inner nuclear membrane and pinch off into the perinuclear space to form a primary enveloped particle. These particles subsequently fuse their envelopes with the outer nuclear membrane and the now cytoplasmic capsids undergo a secondary envelopment in the cytoplasm to form the mature virion. At each envelopment step a complex of proteins termed the tegument is assembled between the capsid and envelope and likely drives the envelopment process. The primary enveloped and mature HSV particles are morphologically quite distinct and are believed to contain quite different arrays of tegument and envelope proteins. However the structure and composition of the perinuclear HSV virus (pnHSV) is poorly defined because to date it has only been possible to examine it by ultrastructural studies. In this application I describe methodology that we have developed to purify pnHSV particles in biochemically useful quantities. We propose to extend our ongoing studies by performing a proteomic analysis of the pnHSV and to test the role of cellular proteins which we have already shown to be present. Furthermore, using the model tegument protein vhs we will dissect the molecular mechanism by which a tegument polypeptide is targeted into the enveloping viral particle. PUBLIC HEALTH RELEVANCE: Herpes simplex viruses (HSV) types 1 and 2 are best known for their role in the generation of oral and genital mucocutaneous lesions, however HSV infection of the central nervous system can also lead to fatal encephalitis, and replication in the eye is a leading cause of blindness in the United States each year. Correct assembly of the HSV particle is critical for the production of an infectious virion, and the spread of disease. This application investigates HSV assembly at the molecular level and will have great therapeutic significance, since it is expected to yield a large and diverse array of new targets for anti-HSV drugs.

描述（由申请方提供）：所有疱疹病毒在感染细胞核中组装其DNA包装衣壳。然后这些衣壳经历了一个生物学上独特的过程：它们发芽进入内核膜，并进入核周空间，形成一个初级包膜颗粒。这些颗粒随后将它们的包膜与外核膜融合，并且现在的细胞质衣壳在细胞质中经历二次沉淀以形成成熟的病毒体。在每一个包被步骤中，被称为被膜的蛋白质复合物在衣壳和包膜之间组装，并可能驱动包被过程。 初级包膜和成熟HSV颗粒在形态上是相当不同的，并且被认为含有相当不同的被膜和包膜蛋白阵列。然而，核周HSV病毒（pnHSV）的结构和组成定义不明确，因为迄今为止，只能通过超微结构研究对其进行检查。在本申请中，我描述了我们已经开发的以生物化学上有用的量纯化pnHSV颗粒的方法。我们建议通过对pnHSV进行蛋白质组学分析来扩展我们正在进行的研究，并测试我们已经证明存在的细胞蛋白的作用。此外，我们将使用模型被膜蛋白vhs来剖析被膜多肽靶向进入包膜病毒颗粒的分子机制。 公共卫生关系：1型和2型单纯疱疹病毒（HSV）最为人所知的是它们在口腔和生殖器粘膜皮肤病变中的作用，然而中枢神经系统的HSV感染也可导致致命的脑炎，并且在眼睛中的复制是美国每年失明的主要原因。HSV颗粒的正确组装对于感染性病毒体的产生和疾病的传播至关重要。该应用在分子水平上研究HSV组装，并将具有巨大的治疗意义，因为它有望产生大量不同的抗HSV药物新靶点。