The DNA Packaging Motor of Bacteriophage Phi29

噬菌体 Phi29 的 DNA 包装马达

基本信息

批准号：
8183408
负责人：
SHELLEY GRIMES
金额：
$ 31.96万
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/8183408
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 Image In Vitro Link Maps Mechanics Methods Modeling Molecular Motors Motion Motor Motor Activity Movement Mutagenesis Mutate Nature Nucleic Acids Organism Pharmaceutical Preparations Process Proteins RNA Binding Research Resolution Roentgen Rays Role Site Structure System Testing Viral Viral Genome Viral Packaging Virus Work biological systems density design flexibility genetic analysis image processing insight macromolecule member molecular recognition nanometer novel operation 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).

双链DNA病毒的DNA包装机制将研究枯草芽孢杆菌噬菌体»29，最有效的体外病毒包装系统已知。使用综合遗传，生化和结构方法，我们将表征蛋白质构象变化和瞬时组装的运动 DNA封装过程中包装电机。包装的机制29将用作类似疱疹病毒中动物病毒包装的模型和腺病毒系统，并帮助寻找新的抗病毒疗法。由于相似性在29 ATPase和其他环易位酶之间，从研究中获得的见解 „ 29包装还将洞悉大分子电动机的基本原理在较高的生物体中起作用。阐明DNA包装的机制：原子 €29 DNA包装电动机的结构将通过X射线拟合获得运动组件的晶体学结构成高分辨率的冷冻图电机复合体（AIM 1）；包装的高分辨率冷冻重建中间体将在DNA包装期间识别移动元素（AIM 2）；和诱变和生化分析将在ATPase中剖析功能残基对于运动功能至关重要的连接器，包括协调和通信，在DNA包装期间（AIM 3）。