The DNA Packaging Motor of Bacteriophage Phi29

噬菌体 Phi29 的 DNA 包装马达

基本信息

批准号：
8337276
负责人：
SHELLEY GRIMES
金额：
$ 31.34万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8337276
关键词：
ATP Hydrolysis ATP phosphohydrolase Address Adenoviruses Amino Acids Animal Viruses Antiviral Therapy Bacillus subtilis Bacteriophages Binding Biochemical Biochemical Genetics Biological Biological Models Capsid Catalysis Cell division Chemicals Chromosome Segregation Communication Complex Cryoelectron Microscopy DNA DNA Binding DNA Packaging Development Disease Double Stranded DNA Virus Electrostatics Elements Eukaryotic Cell Event Family Genetic Genetic Recombination Genome Health Herpesviridae Hydrolysis In Vitro Indium Link Maps Mechanics Modeling Molecular Motors Motion Motor Movement Mutagenesis Mutate Nature Nucleic Acids Organism Pharmaceutical Preparations Process Proteins RNA Binding Research Resolution Roentgen Rays Role Structure System Testing Viral Viral Genome Viral Packaging Virus Work biological systems density design flexibility genetic analysis insight macromolecule member molecular recognition nanometer novel reconstruction self assembly therapeutic target translocase viral DNA

项目摘要

DESCRIPTION (provided by applicant): The mechanism of DNA packaging for double-stranded DNA viruses will be studied in the Bacillus subtilis bacteriophage X29, the most efficient in vitro viral packaging system known. Using an integrated genetic, biochemical and structural approach, we will characterize protein conformational change and movement in the transiently assembled packaging motor during DNA encapsidation. The mechanism of packaging in X29 will serve as a model for animal virus packaging in the analogous herpesvirus and adenovirus systems, and aid in the search for new antiviral therapies. Due to similarities between the X29 ATPase and other ring translocases, insights gained from the study of X29 packaging will also provide insight into the basic principles of macromolecular motor function in higher organisms. To elucidate the mechanism of DNA packaging: an atomic structure of the X29 DNA packaging motor will be obtained by fitting of X-ray crystallographic structures of motor components into high-resolution cryoEM maps of the motor complex (Aim 1); high-resolution cryoEM reconstruction of packaging intermediates will identify mobile elements during DNA packaging (Aim 2); mutagenesis and biochemical analysis will dissect functional residues in the ATPase and connector that are crucial for DNA packaging (Aim 3); and, packaging motors containing chimeric pRNAs will be created to probe the global assembly, operation and communication of the X29 DNA packaging motor (Aim 4).

描述（由申请人提供）：将在枯草芽孢杆菌噬菌体X29中研究双链DNA病毒的DNA包装机制，这是已知的最有效的体外病毒包装系统。使用综合遗传，生化和结构方法，我们将表征DNA封装过程中瞬时组合包装运动中蛋白质构象变化和运动。 X29中包装的机制将作为类似疱疹病毒和腺病毒系统中动物病毒包装的模型，并有助于寻找新的抗病毒疗法。由于X29 ATPase与其他环易位酶之间的相似之处，因此从X29包装研究中获得的见解也将提供对较高生物体中大分子运动功能的基本原理的见解。为了阐明DNA包装的机理：将通过将运动组件的X射线晶体学结构拟合到运动络合物的高分辨率冷冻地图中（AIM 1）来获得X29 DNA包装电机的原子结构；包装中间体的高分辨率冷冻重建将在DNA包装期间识别移动元素（AIM 2）；诱变和生化分析将剖析ATPase和Recontor中对DNA包装至关重要的功能残基（AIM 3）；并且，将创建包含嵌合PRNA的包装电机，以探测X29 DNA包装电机的全局组装，操作和通信（AIM 4）。