The DNA Packaging Motor of Bacteriophage Phi29

噬菌体 Phi29 的 DNA 包装马达

基本信息

批准号：
8726431
负责人：
SHELLEY GRIMES
金额：
$ 30.06万
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 2016-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8726431
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

The mechanism of DNA packaging for double-stranded DNA viruses will be studied in the Bacillus subtilis bacteriophage ¿29, 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 ¿29 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 ¿29 ATPase and other ring translocases, insights gained from the study of ¿29 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 ¿29 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); and mutagenesis and biochemical analysis will dissect functional residues in the ATPase and connector that are crucial for motor function, including coordination and communication, during DNA packaging (Aim 3).

将研究双链DNA病毒的DNA包装机制枯草芽孢杆菌噬菌体 ¿ 29、最高效的体外病毒包装系统我们将利用综合遗传、生化和结构方法。表征瞬时组装的蛋白质构象变化和运动 DNA 衣壳化过程中的包装电机 ¿ 中的包装机制。 29 会作为类似疱疹病毒中动物病毒包装的模型，腺病毒系统，并有助于寻找新的抗病毒疗法。之间 ¿ 29 ATP 酶和其他环转位酶，从研究中获得的见解 ¿ 29包装还将深入了解高分子电机的基本原理阐明 DNA 包装的机制：原子。的结构 ¿通过X射线拟合获得29个DNA包装马达将电机组件的晶体结构转化为高分辨率冷冻电镜图运动复合体（目标 1）；包装的高分辨率冷冻电镜重建中间体将在 DNA 包装过程中识别移动元件（目标 2）；诱变和生化分析将剖析 ATP 酶中的功能残基，对运动功能至关重要的连接器，包括协调和沟通， DNA 包装期间（目标 3）。