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(申请人提供)：所有疱疹病毒都在感染的细胞核中组装DNA包装的衣壳。然后，这些衣壳经历了一个生物学上独一无二的过程：它们发芽进入内核膜，然后掐断进入核周空间，形成一个初级被包裹的粒子。这些颗粒随后将其包膜与外核膜融合，现在的细胞质衣壳在细胞质中经历二次包膜，形成成熟的病毒粒子。在每个包膜步骤中，一个被称为被膜的蛋白质复合体被组装在衣壳和包膜之间，并可能驱动包膜过程。原始的包膜和成熟的单纯疱疹病毒颗粒在形态上非常不同，被认为含有完全不同的被膜和包膜蛋白阵列。然而，核周单纯疱疹病毒(PnHSV)的结构和组成尚不清楚，因为到目前为止，只能通过超微结构研究来检查它。在本申请中，我描述了我们开发的以生物化学有用的数量提纯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质的作用。此外，使用被膜蛋白VHS的模型，我们将剖析被膜多肽被靶向包膜病毒颗粒的分子机制。 公共卫生相关性：单纯疱疹病毒(HSV)1型和2型因其在口腔和生殖器粘膜皮肤损害中的作用而闻名，然而HSV感染中枢神经系统也可导致致命的脑炎，在美国，眼睛复制是每年导致失明的主要原因。HSV粒子的正确组装对于产生具有感染性的病毒粒子和疾病传播至关重要。这项应用在分子水平上研究HSV组装，将具有重大的治疗意义，因为它有望产生大量和多样化的抗HSV药物的新靶点。