ROTAVIRUS, NORWALK VIRUS, AND ORTHOREOVIRUSES

轮状病毒、NORWALK 病毒和正病毒

• 批准号: 8361057
• 负责人: Bidadi Venkataram Prasad
• 金额: $ 2.45万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361057
• 关键词: Ammonium Chloride; Animals; Antibodies; Antiviral Agents; BCAR3 gene; Biochemical Genetics; Caliber; Caliciviridae; Calicivirus; Capsid; Cells; Child; Collaborations; Complex; Diarrhea; Electron Microscopy; Epidemic; Exhibits; Funding; Gastroenteritis; Genetic Transcription; Genome; Goals; Grant; Human; Image; In Vitro; Infection; Influenza A Virus, H5N1 Subtype; Interferon Type I; Kinetics; Life; Medical; Membrane; Morphogenesis; National Center for Research Resources; Nature; Norovirus; Norwalk virus; Orthoreoviruses; PTPN11 gene; Pathogenicity; Penetration; Principal Investigator; Proteins; Proteolysis; Reoviridae Infections; Reovirus; Research; Research Infrastructure; Research Project Grants; Resolution; Resources; Rice; Rotavirus; Source; Structure; Structure-Activity Relationship; System; United States National Institutes of Health; Virulence; Virus; Work; base; cost; epizootic; global health; helicase; image processing; influenza virus INS1 protein; influenzavirus; inhibitor/antagonist; insight; interest; macromolecule; member; mutant; nanometer; nucleoside triphosphatase; particle; pathogen; protein function; receptor; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Major thrust of our research projects is to understand structure-function relationships in medically important viruses such as rotavirus (NIH AI36040), caliciviruses (NIH P01 AI57788-01 in collaboration with Dr. M.K. Estes), orthoreoviruses (NIH RO1 AI32539 in collaboration with Dr. T. Dermody), and influenza viruses (in collaboration with Dr. Andrew Rice, grant submission in progress). We have been using NCRR funded NCMI facility over years, and we will continue to use this excellent facility. The world-class high resolution imaging and image processing facilities are excellently suited for our projects. Rotavirus: Rotavirus is the major cause of severe, life-threatening gastroenteritis in young children and animals. Rotaviruses are large (1000 ¿ ), complex, icosahedral assemblies. This virus has been the subject of extensive biochemical, genetic and structural studies because of its medical relevance, intriguing structural complexity, and unique strategies of replication and morphogenesis. We will continue further understanding of the structural basis of rotavirus cell entry, endogenous transcription, viroplasm formation, genome replication and packaging at sub-nanometer resolution using the NCMI cryo-EM facility. Our focus will be particularly on human strains. While some of these studies involve rotavirus particles (1000 ¿ diameter), others that are related to viroplasm formation, genome replication/packaging involving rotavirus non-structural proteins such as NSP2, NSP5, VP1, VP2 and NSP6. In infected cells, these proteins work in concert during viroplasm formation, genome replication/encapsidation. To obtain structural insights into how these rotavirus proteins function, high-resolution cryo-EM analysis is perhaps the only option. Caliciviruses: Caliciviruses, grouped into four genera, are important human and veterinary pathogens with a potential for zoonosis2. In these ssRNA viruses, capsid related functions such as assembly, antigenicity, and receptor interactions are encoded in a single protein that forms an icosahedral capsid. We are interested in obtaining structural information about members of the Caliciviridae. We will continue to use NCMI facility to investigate calicivirus-receptor, and calicivirus-antibody interactions. In addition, we plan to use these resources available at NCMI for characterizing norovirus p41 protein that exhibits helicase/NTPase activities. Noroviruses are causative agents of epidemic diarrhea in humans 3. Orthoreoviruses: The goal of the proposed research is to determine mechanisms of non- enveloped virus internalization, disassembly, and membrane penetration using mammalian reovirus as an experimental system. We will continue to use NCMI facilities to further investigate structural basis for reovirus disassembly and membrane penetration by determining structures of sequential reovirus disassembly intermediates, and that of reovirus mutants selected during persistent reovirus infection (PI viruses), which demonstrate enhanced in vitro kinetics of disassembly and grow in the presence of inhibitors of endosomal acidification (ammonium chloride)4 and proteolysis. Influenza virus NS1 protein: Recent emergence of highly pathogenic avian (H5N1) influenza viruses, their epizootic and panzootic nature, and their association with lethal human infections have raised significant global health concerns. Several studies have underscored the importance of non-structural protein NS1 in the increased pathogenicity and virulence of these strains. NS1, is an antagonist of antiviral type-I interferon (IFN) response in the host.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们的研究项目的主要目的是了解结构与功能的关系， 医学上重要的病毒如轮状病毒（NIH AI 36040）、杯状病毒（NIH P01 AI 57788 -01与M.K.博士合作Estes）、正呼肠孤病毒（NIH R 01 AI 32539， 与T博士合作Dermody）和流感病毒（与Andrew博士合作 Rice，grant submission in progress）. 我们一直在使用NCRR资助的NCMI设施， 多年来，我们将继续使用这一出色的设施。 世界级的高分辨率 成像和图像处理设备非常适合我们的项目。 轮状病毒：轮状病毒是引起严重的、危及生命的年轻人胃肠炎的主要原因 孩子和动物。 轮状病毒是大型（1000 <$）、复杂的二十面体组装体。 这种病毒一直是广泛的生化、遗传和结构研究的对象 因为它的医学相关性，有趣的结构复杂性，以及独特的策略， 复制和形态发生。 我们将继续进一步了解结构 轮状病毒进入细胞的基础，内源性转录，病毒质形成，基因组 复制和包装在亚纳米分辨率使用NCMI冷冻EM设施。 我们将特别关注人类菌株。 虽然其中一些研究涉及 轮状病毒颗粒（直径1000 <$），其他与病毒质形成有关的颗粒， 涉及轮状病毒非结构蛋白如NSP 2的基因组复制/包装， NSP 5、VP 1、VP 2和NSP 6。 在受感染的细胞中，这些蛋白质协同工作， 病毒质形成、基因组复制/降解。 为了获得结构上的洞察力， 这些轮状病毒蛋白的功能，高分辨率冷冻电镜分析可能是 唯一的选择 杯状病毒：杯状病毒分为四个属，是人类和兽医的重要病毒。 具有人畜共患病潜力的病原体2. 在这些ssRNA病毒中，衣壳相关功能 例如装配、抗原性和受体相互作用在单个蛋白质中编码 形成二十面体衣壳 我们感兴趣的是获得结构信息 关于杯状病毒科成员的信息 我们将继续使用NCMI设施进行调查 杯状病毒-受体和杯状病毒-抗体相互作用。 此外，我们计划利用这些 NCMI提供的用于表征诺如病毒p41蛋白的资源， 解旋酶/NTR酶活性。 诺如病毒是我国流行性腹泻的病原体， 人类3. 正呼肠孤病毒：拟议研究的目标是确定非- 包膜病毒的内化、分解和膜穿透， 哺乳动物呼肠孤病毒作为实验系统。我们将继续使用NCMI设施， 进一步研究呼肠孤病毒分解和膜穿透的结构基础， 确定顺序呼肠孤病毒拆卸中间体的结构，以及 在持续性呼肠孤病毒感染（PI病毒）期间选择的呼肠孤病毒突变体， 证明了在存在以下物质的情况下增强的体外分解和生长动力学 内体酸化（氯化铵）4和蛋白水解的抑制剂。 流感病毒NS 1蛋白：最近出现的高致病性禽流感（H5 N1） 流感病毒，它们的流行性和泛动物性， 人类感染引起了严重的全球健康问题。几项研究 强调了非结构蛋白NS 1在增加致病性中的重要性 这些菌株的毒性。 NS 1是抗病毒I型干扰素（IFN）的拮抗剂， 在主持人的回答。