Molecular analysis of astrovirus capsid assembly

星状病毒衣壳组装的分子分析

• 批准号: 6809605
• 负责人: NEEL KUMAR KRISHNA
• 金额: $ 20.46万
• 依托单位: EASTERN VIRGINIA MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6809605
• 关键词: Astrovirus; Baculoviridae; capsid; chimeric proteins; gastroenteritis; gastrointestinal infection; gene expression; gene mutation; genetic manipulation; life cycle; molecular cloning; molecular site; mutant; polymerase chain reaction; protein sequence; protein structure function; virion; virus RNA; virus assembly; virus genetics; virus protein

DESCRIPTION (provided by applicant): Coat proteins of non-enveloped, icosahedral viruses must perform a variety of functions during the virus life cycle, such as assembly of the coat protein subunits into a closed shell, specific encapsidation of the viral nucleic acid, maturation of the capsid, interaction with host receptors and disassembly to deliver the genetic information into the newly-infected cell. A thorough understanding of the multiple capsid properties at the molecular level is required in order to identify potent targets for antiviral therapy and the prevention of viral disease. The systems we have chosen for study are the astroviruses, a family of icosahedral, single-stranded RNA virus that is a causative agent of gastroenteritis in both humans and animals. Very little is known about what regions of the coat protein contribute to the diverse capsid functions. The objective of this application is to test whether novel structural predictions we have made based on the coat protein sequence of the human astroviruses are exhibited in biological experiments. We hypothesize that the assembly and RNA packaging functions of the astrovirus coat protein constitute an individual, modular domain that is distinct from the determinants required for receptor binding and internalization. Our specific aims are: (1) To test whether functional predictions of regions of the coat protein required for virion assembly, particle maturation, and RNA encapsidation are exhibited in biological experiments, by characterizing the phenotypes of a series of mutations. (2) To test the modularity of the coat protein "assembly domain" by constructing chimeric coat protein molecules between different serotypes and species of the Astroviridae. The rationale for these studies is that insight into the molecular mechanisms of astrovirus assembly, nucleic acid packaging, maturation and tropism will inform us of unique and shared properties of astrovirus coat proteins. In addition, the potential to engineer chimeric astrovirus particles that display specific epitopes on the surface of the particle could have practical applications such as generation of particle-based vaccines. Chimeric astrovirus particles displaying surface antigens of related gastroenteritis virus such as norovirus, a Category B bioterrorism agent, could be developed as a polyvalent, preventative vaccine. In the long term, we expect that this work will add not only to the understanding of astrovirus particle biology, but also lead to the development of therapeutic drugs or vaccines to combat viral gastroenteritis in humans.

描述（由申请人提供）：非发育的二十面体病毒的涂层蛋白必须在病毒生命周期中执行多种功能，例如将涂料蛋白亚基组装成封闭的壳，特定的病毒核酸的封装，成熟在衣壳中，与宿主受体的相互作用和拆卸，以将遗传信息传递到新感染的细胞中。为了确定抗病毒疗法和预防病毒疾病的有效靶标，需要对分子水平上多重衣壳特性进行透彻的了解。我们选择的研究系统是Astroviruses，它是一个二十面体，单链RNA病毒的家族，是人类和动物的胃肠炎的致病药物。关于大衣蛋白的哪些区域对不同的衣壳功能的贡献，知之甚少。该应用的目的是测试我们在生物学实验中表现出我们基于人类星座病毒的外套蛋白序列做出的新的结构预测。我们假设，星形病毒外套蛋白的组装和RNA包装函数构成了一个单独的模块化结构域，该结构域与受体结合和内在化所需的决定因素不同。我们的具体目的是：（1）通过表征一系列突变的表型来测试在生物实验中表现出病毒体组装，颗粒成熟和RNA封装所需的外套蛋白区域的功能预测。 （2）通过在不同的血清型和Astroviridae物种之间构造嵌合外套蛋白分子来测试外套蛋白“装配结构域”的模块化。这些研究的基本原理是，洞悉赤道病毒组装，核酸包装，成熟和tropism的分子机制将为我们告知我们的星座大衣蛋白的独特和共享特性。此外，在粒子表面上显示特定表位的嵌合式星形病毒颗粒的潜力可能具有实用的应用，例如产生颗粒的疫苗。可以开发出一种多价，预防性疫苗的B类生物恐怖剂的嵌合星体颗粒，这些颗粒显示出相关胃肠炎病毒的表面抗原，例如B类生物恐怖剂。从长远来看，我们预计这项工作不仅会增加对星座颗粒生物学的理解，而且还会导致治疗药物或疫苗的发展以对抗人类的病毒胃肠炎。