ROTAVIRUS AND NORWALK VIRUS

轮状病毒和诺沃克病毒

• 批准号: 8168527
• 负责人: Bidadi Venkataram Prasad
• 金额: $ 2.15万
• 依托单位: BAYLOR 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: 2010-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168527
• 关键词: Ammonium Chloride; Animals; Antibodies; Antiviral Agents; BCAR3 gene; Biochemical Genetics; Caliber; Caliciviridae; Calicivirus; Capsid; Cells; Child; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; Diarrhea; Epidemic; Exhibits; Funding; Gastroenteritis; Genetic Transcription; Genome; Goals; Grant; Human; Image; In Vitro; Infection; Influenza A Virus, H5N1 Subtype; Institution; Interferon Type I; Kinetics; Life; Medical; Membrane; Morphogenesis; National Center for Research Resources; Nature; Norovirus; Norwalk virus; Orthoreoviruses; PTPN11 gene; Pathogenicity; Penetration; Proteins; Proteolysis; Reoviridae Infections; Reovirus; Research; Research Personnel; Research Project Grants; Resolution; Resources; Rice; Rotavirus; Source; Structural Protein; Structure; Structure-Activity Relationship; System; United States National Institutes of Health; Virulence; Virus; Work; base; epizootic; global health; helicase; image processing; influenza virus INS1 protein; influenzavirus; inhibitor/antagonist; insight; interest; 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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们研究项目的主要目标是了解医学上重要的病毒的结构-功能关系，例如轮状病毒（NIH AI36040）、杯状病毒（与 M.K. Estes 博士合作的 NIH P01 AI57788-01）、正呼肠孤病毒（与 T. Dermody 博士合作的 NIH RO1 AI32539）和流感病毒（与 Andrew 博士合作） 赖斯，正在提交赠款）。 多年来，我们一直在使用 NCRR 资助的 NCMI 设施，我们将继续使用这一出色的设施。 世界一流的高分辨率成像和图像处理设施非常适合我们的项目。 轮状病毒：轮状病毒是幼儿和动物患严重、危及生命的胃肠炎的主要原因。 轮状病毒是大型（1000 ¿）、复杂的二十面体组装体。 由于其医学相关性、令人感兴趣的结构复杂性以及独特的复制和形态发生策略，该病毒已成为广泛的生化、遗传和结构研究的主题。 我们将继续使用 NCMI 冷冻电镜设备以亚纳米分辨率进一步了解轮状病毒细胞进入、内源转录、病毒质形成、基因组复制和包装的结构基础。 我们的重点将特别放在人类菌株上。 虽然其中一些研究涉及轮状病毒颗粒（直径为 1000 ¿），但其他研究与病毒质形成、基因组复制/包装有关，涉及轮状病毒非结构蛋白，如 NSP2、NSP5、VP1、VP2 和 NSP6。 在受感染的细胞中，这些蛋白质在病毒质形成、基因组复制/衣壳化过程中协同工作。 为了深入了解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，是重要的人类和兽医病原体，具有人畜共患病的潜力2。 在这些 ssRNA 病毒中，衣壳相关功能（例如组装、抗原性和受体相互作用）被编码在形成二十面体衣壳的单个蛋白质中。 我们有兴趣获得有关杯状病毒科成员的结构信息。 我们将继续使用 NCMI 设施来研究杯状病毒-受体和杯状病毒-抗体相互作用。 此外，我们计划利用 NCMI 提供的这些资源来表征具有解旋酶/NTPase 活性的诺如病毒 p41 蛋白。 诺如病毒是人类流行性腹泻的病原体3。 正呼肠孤病毒：本研究的目标是使用哺乳动物呼肠孤病毒作为实验系统来确定无包膜病毒内化、分解和膜渗透的机制。我们将继续使用 NCMI 设施，通过确定连续呼肠孤病毒分解中间体的结构以及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的结构，进一步研究呼肠孤病毒分解和膜渗透的结构基础，这些突变体证明在内体酸化抑制剂（铵）存在下增强了体外分解和生长的动力学。 氯化物）4 和蛋白水解。 流感病毒 NS1 蛋白：最近出现的高致病性禽 (H5N1) 流感病毒、其流行性和泛动物性及其与致命人类感染的关联引起了全球健康问题。多项研究强调了非结构蛋白 NS1 在增加这些菌株的致病性和毒力中的重要性。 NS1 是宿主抗病毒 I 型干扰素 (IFN) 反应的拮抗剂。