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: 8050596
• 负责人: DUNCAN W. WILSON
• 金额: $ 56.54万
• 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/8050596
• 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（由申请人提供）：所有疱疹病毒在感染的细胞核中组装了其DNA包装的衣壳。然后，这些衣壳经历了生物学独特的过程：它们进入内部核膜，并捏入核周空间以形成主要的包裹粒子。这些颗粒随后将其信封与外核膜和现在的细胞质衣壳融合在一起，在细胞质中经历次级包膜，形成成熟的病毒粒子。在每个包络步骤中，称为Tegument的蛋白质复合物是在衣壳和包膜之间组装的，并可能驱动了信封过程。 主要的包围和成熟的HSV颗粒在形态上是完全不同的，并且被认为包含截然不同的tegument和包膜蛋白。但是，核周HSV病毒（PNHSV）的结构和组成的定义很差，因为迄今为止，只有通过超微结构研究才有可能检查它。在此应用程序中，我描述了我们已经开发出来纯化PNHSV颗粒在生化有用数量中的方法。我们建议通过对PNHSV进行蛋白质组学分析并测试我们已经证明存在的细胞蛋白的作用来扩展我们正在进行的研究。此外，使用模型Tegument蛋白VHS，我们将剖析将tegument多肽靶向被包围的病毒颗粒的分子机制。 公共卫生相关性：单纯疱疹病毒（HSV）类型1和2以其在口腔和生殖器粘膜损伤中的作用而闻名，但是中枢神经系统的HSV感染也会导致致命的脑炎，并在眼中复制。每年是美国失明的主要原因。 HSV颗粒的正确组装对于产生感染性病毒体和疾病的传播至关重要。该应用在分子水平上研究了HSV组装，并且具有很高的治疗意义，因为预计它将为抗HSV药物产生大量而多样的新靶标。