Multisubunit viral ATPases that couple ATP-hydrolysis to genome translocation

将 ATP 水解与基因组易位耦合的多亚基病毒 ATP 酶

• 批准号: 8238803
• 负责人: Gino Cingolani
• 金额: $ 29.45万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8238803
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Architecture; Bacteriophage P22; Binding; Biochemical; Biology; Burn injury; Capsid; Chemicals; Chemistry; Complex; Couples; DNA; DNA Viruses; Data; Double Stranded DNA Virus; Enzymes; Escherichia coli; F1-ATPase; GTPase-Activating Proteins; Genome; Goals; Grant; Herpesviridae; Holoenzymes; Human Virus; Laboratories; Life; Literature; Macromolecular Complexes; Methods; Mitochondria; Molecular; Molecular Conformation; Molecular Machines; Motor; Nature; Process; Proteins; Reaction; Recruitment Activity; Research; Resolution; Role; Structure; Substrate Specificity; Techniques; Testing; Translations; Viral; Viral Genome; Viral Packaging; Work; bacteriophage P22 portal protein; base; novel; stoichiometry; terminase; vacuolar H+-ATPase; viral DNA

DESCRIPTION (provided by applicant): Packaging of viral genomes is a fundamental process in biology. In many DNA viruses this reaction is powered by a large (M.W. ~1.5MDa) genome-packaging motor, which is formed by a terminase holoenzyme assembled to a dodecameric portal protein. This macromolecular complex functions like a chemical motor, which hydrolyzes ATP to translocate a copy of the viral genome inside a preformed capsid, at rates as high as 2000 bases per second. In addition to being the fastest and most powerful engine in nature, the genome-packaging motor is also an intriguing molecular machine, which is poorly characterized both in structure and catalytic mechanisms. In this grant, we will use a combination of crystallographic and biochemical techniques to characterize the structure of the bacteriophage P22 genome-packaging motor. Building upon the structure of P22 portal protein, which was recently determined in my laboratory, our work will focus on the chemistry of P22 terminase that is a functional ATPase in the motor. The structural characterization of P22 terminase in complex with portal protein will provide a structural framework to decipher how, within the packaging motor, terminase couples ATP hydrolysis to translocation of viral genomes. In addition, since terminases are highly conserved in herpesviruses, the work proposed in this grant will also provide a logical framework to begin structural characterization of packaging motors in pathogenic human viruses. Specific aims of our work are: 1.) to determine the structure of the P22 terminase holoenzyme assembled at the portal protein vertex; 2.) to define how small terminase subunit stimulates large terminase ATPase activity during genome-packaging. PUBLIC HEALTH RELEVANCE: Viral genome-translocating motors are multisubunit ATPases that burn ATP to power packaging of viral genomes inside a preformed capsid. In this grant, we propose to study the structure and activity of the bacteriophage P22 genome-translocating motor. Our work will determine how one of smallest motors in nature couples chemical energy to genome translocation.

描述（由申请人提供）：病毒基因组的包装是生物学的基本过程。在许多DNA病毒中，该反应由大型（M.W. 〜1.5MDA）基因组包装运动供电，该电动机由组装到DeeteCameric Portal蛋白的末端酶全酶形成。该大分子复合物的作用像化学电动机，该化学电动机水解ATP以每秒高达2000碱基的速率将病毒基因组的副本转移到了预制的衣壳内。除了是最快，最强大的发动机外，基因组包装电动机还是一款有趣的分子机器，在结构和催化机制中的特征都很差。在这笔赠款中，我们将使用晶体学和生化技术的组合来表征噬菌体P22基因组包装运动的结构。在我的实验室最近确定的P22门户蛋白的结构的基础上，我们的工作将集中在P22末端酶的化学上，该化学是电动机中功能性ATPase。 p22末端酶与门户蛋白的复合物中的结构表征将提供一个结构框架，以破译包装运动中的末端酶ATP水解如何水解为病毒基因组的易位。此外，由于末端酶在疱疹病毒中是高度保守的，因此该赠款中提出的工作还将提供一个逻辑框架，以开始对病原体病毒中包装电机的结构表征。我们工作的具体目的是：1。）确定在门户蛋白顶点组装的p22末端酶全酶的结构； 2.）定义小末端亚基如何刺激基因组包装过程中的大末端ATPase活性。 公共卫生相关性：病毒基因组转化电动机是多生育ATPases，可将ATP燃烧到预先形成的衣壳内的病毒基因组的功率包装。在这笔赠款中，我们建议研究噬菌体p22基因组转移运动的结构和活性。我们的工作将确定自然界中最小的电动机之一化学能量到基因组易位。