The Molecular Structure of the RNA Polymerase Portal of a Bacteriophage-Reovirus

噬菌体呼肠孤病毒 RNA 聚合酶门户的分子结构

基本信息

批准号：
8269906
负责人：
Paul Jeffrey Gottlieb
金额：
$ 34.3万
依托单位：
CITY COLLEGE OF NEW YORK
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8269906
关键词：
Adolescent Antibodies Antiviral Agents Arts Bacteria Bacteriophages Biological Assay Capsid Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Characteristics Childhood Cities Clinical Collaborations Communities Complex Computer software Cryoelectron Microscopy Cystovirus DNA DNA-Directed RNA Polymerase Data Developing Countries Development Diarrhea Doctor of Medicine Doctor of Philosophy Double-Stranded RNA Drug Delivery Systems Drug Formulations Education Educational Curriculum Educational workshop Electron Microscopy Electrons Elements Emerging Communicable Diseases Family Fluorescence Foundations Funding Gastroenteritis Genes Genetic Engineering Genetic Transcription Genome Genomic Segment Goals Grant Hospitalization Human Hydration status Image Immunology Indium Infection Influenza Institutes Institution Knowledge Laboratories Laboratory Personnel Laboratory Research Lasers Learning Light-Scattering Spectroscopy Medical Medical Students Microbiology Minority Modeling Molecular Biology Molecular Machines Molecular Models Molecular Structure Monoclonal Antibodies Morbidity - disease rate Motor New York New York City Nucleocapsid Nucleotides Optical Methods Optics Paper Pharmacologic Substance Polymerase Positioning Attribute Postdoctoral Fellow Preparation Primary Care Physician Process Proteins Publications Publishing Qualifying RNA RNA chemical synthesis RNA replication Recombinants Reoviridae Reovirus Reproduction Research Research Infrastructure Research Personnel Research Project Grants Research Proposals Resistance Resources Role Rotavirus Schools Science Shapes Side Spectrum Analysis Staining method Stains Structural Models Structure Students System Technology Testing Tomogram Training Transcript United States United States National Institutes of Health Vaccines Viral Viral Packaging Viral Proteins Virus Virus Diseases Work Writing base career clinically significant college design doctoral student genetic regulatory protein graduate student in vitro Assay innovation medical schools medically underserved meetings microbial molecular modeling mutant nucleoside triphosphatase particle pathogen professor programs public health relevance reconstruction structural biology symposium technology/technique tomography tool viral RNA

项目摘要

DESCRIPTION (provided by applicant): Our study is directed towards understanding the interactions that exist among the proteins that constitute the RNA polymerase portal of the cystoviruses. This molecular machine selects, packages, and replicates the viral RNA genome segments. One portal is located on each vertex of the 12-sided polymerase complex (PX) a structure that forms the viral core. This bacteriophage family constitutes a unique bacterial reovirus that serves as a simple and important molecular model for their clinically significant cousins, the Reoviridae. Our overall objective is to establish a detailed structural model for the reoviruses portal apparatus using cryo-electron microscopy (cryo-EM). We will then better understand how they replicate and package their double-stranded RNA (dsRNA) genomes. This knowledge is of critical importance in the design of antiviral pharmaceuticals for pathogenic reoviruses, in particular rotavirus, a significant cause of childhood morbidity in third world countries. My developmental and career objective is to employ a qualified research team that studies the molecular biology and structure of clinically significant viruses at the City College of New York (CCNY) Medical School. The cystovirus model offers an excellent introductory virus project that utilizes the resources for molecular and structural biology recently established at the CCNY campus in particular the New York State Foundation for Science, Technology and Innovation (NYSTAR) funded New York Structural Biology Center (NYSBC). The Gottlieb laboratory personnel have utilized the hardware and software at this new facility and have published the center's first electron-cryo- tomography paper. We are completing a second electron cryo-microscopy paper that constitutes the preliminary study for this proposal. We have recently established a new optical method to examine the dynamic conformational shape changes that occur in viral proteins during the infection process. This research is performed in collaboration with physicists at the Institute for Ultrafast Spectroscopy and Lasers (IUSL) on the CCNY campus. We have included an aim in the research plan directed towards perfecting our optical assays for the analysis of biochemically active cystovirus PX. The PX is known to radically expand during viral RNA packaging and replication. We believe that this portion of our research proposal fits well within the goals of the current project. We anticipate that the optical qualities of viral proteins that we discover are applicable to human pathogens, in particular influenza. PUBLIC HEALTH RELEVANCE: Group A rotaviruses, as noted by the Centers for Disease Control (CDC), are the major cause of juvenile diarrhea leading to several thousand hospitalizations annually in the United States and close to a million deaths worldwide. There is no antiviral formulation to treat this illness and the current vaccine is not 100% effective. Our research utilizes cystoviruses, a type of rotavirus, which only replicate in bacteria cells. This is a system that we can study in great detail to understand the mechanisms of virus reproduction, and provide information critical to the development of anti-rotavirus compounds.

描述（由申请人提供）：我们的研究旨在理解构成猫病毒的RNA聚合酶门户的蛋白质之间存在的相互作用。该分子机选择，包装并复制病毒RNA基因组段。一个门户位于12面聚合酶复合物（PX）的每个顶点上，形成病毒核的结构。这种噬菌体家族构成了独特的细菌丙卷病毒，它是其临床意义的表兄弟Reoviridae的简单而重要的分子模型。我们的总体目的是使用低温电子显微镜（Cryo-EM）建立一个详细的葡萄病毒门户设备的结构模型。然后，我们将更好地理解它们如何复制和包装双链RNA（DSRNA）基因组。这种知识对于致病性静脉病毒的抗病毒药物的设计至关重要，尤其是轮状病毒，这是第三世界国家儿童发病率的重要原因。我的发展和职业目标是雇用一个合格的研究团队，研究纽约市学院（CCNY）医学院的临床意义病毒的分子生物学和结构。膀胱病毒模型提供了一个出色的入门病毒项目，该项目利用最近在CCNY校园建立的分子和结构生物学资源，尤其是纽约州科学，技术与创新基金会（NYSTAR）资助的纽约结构生物学中心（NYSBC）。 Gottlieb实验室人员在此新设施上使用了硬件和软件，并发表了该中心的第一张电子透明纸。我们正在完成第二份电子冷冻微镜纸，该纸构成了该提案的初步研究。我们最近建立了一种新的光学方法，以检查感染过程中病毒蛋白中发生的动态构象形状变化。这项研究是与CCNY校园的超快光谱和激光研究所（IUSL）合作进行的。我们已经将目的包括在研究计划中，该计划旨在完善我们的光学测定法，以分析生化活性膀胱病毒PX。已知PX在病毒RNA包装和复制过程中从根本上扩展。我们认为，我们的研究建议的这一部分非常适合当前项目的目标。我们预计我们发现的病毒蛋白的光学品质适用于人类病原体，尤其是流感。公共卫生相关性：疾病控制中心（CDC）指出的轮状病毒组是少年腹泻的主要原因，导致美国每年在美国进行数千次住院，全球近一百万人死亡。没有用于治疗这种疾病的抗病毒剂，目前的疫苗也不有效。我们的研究利用了半肠病毒，一种轮状病毒，仅在细菌细胞中复制。这是我们可以详细研究以了解病毒繁殖机制的系统，并提供对抗鼠病毒化合物发展至关重要的信息。