Probing the Universality and Function of Single dsDNA Viral Packaging Motors

探讨单 dsDNA 病毒包装马达的通用性和功能

• 批准号: 7561077
• 负责人: james m tsay
• 金额: $ 5.01万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7561077
• 关键词: Adenoviruses; Antiviral Agents; Bacteriophages; Biochemical; Biological Assay; Chemicals; DNA; DNA Packaging; Dependence; Goals; Herpesviridae; Measurement; Measures; Molecular Motors; Motor; Operating System; Poxviridae; Proteins; Public Health; Research; Role; System; Techniques; Testing; Viral Packaging; Virus; Work; design; information gathering; interest; laser tweezer; research study; viral DNA

DESCRIPTION (provided by applicant): The goals of this proposal are to help bring a better understanding of how viral packaging motors function and to find out the role of the components critical to their function. Using single virus and DNA manipulation techniques, we have the opportunity to extract an unprecedented amount of detailed information on how these systems operate. Furthermore, by exploring different viral packaging systems we can test whether certain classes of viruses all work using the same mechanisms. First, an experimental technique will be developed to investigate how lambda viruses package DNA. Second, the detailed mechanism in which the motor can take chemical energy and convert it to moving DNA will be investigated. Finally, the effect of critical proteins to the survival of this virus on its ability to package DNA will be explored. The fundamental information gathered from these experiments may aid the rational design of antiviral drugs. Specific Aims: 1. The universality of viral DNA packaging motors will be explored by developing a new optical tweezers assay for studying DNA packaging in bacteriophage A. Motor force, DNA translocation measurements, and their dependence on biochemical conditions will be extended to A. 2. The mechanochemistry of the lambda viral packaging motor will be elucidated by measuring ATP, ADP, and Pi dependence, pause distributions, and force measurements on packaging. 3. The specific roles of gpD and gpFI, proteins critical for lambda viral packaging, will be investigated. This research will be of interest to public health because the system that will be studied is expected to have similarities to relevant mammalian viruses such as herpesviruses, adenoviruses, and poxviruses. They are thought to share a common packaging mechanism, presumably powered by a similar molecular motor. By understanding the detailed mechanochemistry as well as the additional proteins necessary for the survival of these viruses, we may envision new strategies to regulate their infectivity.

描述（由申请人提供）：该提案的目标是帮助更好地了解病毒包装马达的功能，并找出对其功能至关重要的组件的作用。使用单一病毒和 DNA 操作技术，我们有机会提取有关这些系统如何运作的前所未有的详细信息。此外，通过探索不同的病毒包装系统，我们可以测试某些类别的病毒是否都使用相同的机制发挥作用。首先，将开发一种实验技术来研究 lambda 病毒如何包装 DNA。其次，将研究电机吸收化学能并将其转化为移动 DNA 的详细机制。最后，我们将探讨该病毒的生存关键蛋白对其包装 DNA 能力的影响。从这些实验中收集的基本信息可能有助于抗病毒药物的合理设计。 具体目标： 1. 将通过开发一种用于研究噬菌体 A 中 DNA 包装的新型光镊测定来探索病毒 DNA 包装马达的普遍性。马达力、DNA 易位测量及其对生化条件的依赖性将扩展到 A。 2. 将通过测量 ATP、ADP 和 Pi 依赖性、暂停分布和包装上的力测量来阐明 lambda 病毒包装马达的机械化学。 3. 将研究对 lambda 病毒包装至关重要的蛋白质 gpD 和 gpFI 的具体作用。 这项研究将引起公共卫生的兴趣，因为将要研究的系统预计与相关哺乳动物病毒（如疱疹病毒、腺病毒和痘病毒）有相似之处。人们认为它们具有共同的包装机制，可能由类似的分子马达提供动力。通过了解详细的机械化学以及这些病毒生存所需的其他蛋白质，我们可以设想调节其感染性的新策略。