Structural Analysis of Virulence Determinants and Genomic DNA Packaging Constraints in a ssDNA Virus

单链 DNA 病毒毒力决定因素和基因组 DNA 包装限制的结构分析

• 批准号: 0718948
• 负责人: Mavis Agbandje-McKenna
• 金额: $ 51.18万
• 依托单位: University of Florida
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0718948&HistoricalAwards=false
• 关键词: Structural; Analysis; Virulence; Determinants; Genomic

The three-dimensional (3D) structure of a macromolecule is assembled to enable functional utilization. This 3D structure, however, once conformed, is capable of transitions and/or re-arrangements, as required, to enable essential functional interactions. The consecutive macromolecular interactions that define the life-cycle of viruses during host-cell infection represent excellent examples of the high fidelity of functional utilization of macromolecular structure. However, despite much research on viral life cycles, there is limited 3D structural information on the facilitating capsid-host interactions, structural capsid transitions, and capsid-genome interactions. This project will utilize naturally-occurring variants and emerging virulent strains of the Minute Virus of Mice (MVM), a member of the ssDNA Parvoviridae, as a viral model to decipher the structural requirements for successful host-cell receptor recognition, cellular trafficking and capsid maturation triggered by genomic interaction. This model system is also ideal for studies of virus strains that are similar to one another yet have pronounced differences in their host-range, tissue tropism, and pathogenic outcomes of infection due to minor structural variations or amino-acid differences on their capsid viral proteins which modify required infectious interactions. These variations often confer altered viral host-cell interactions, including infectious receptor utilization, which is common in emerging viruses. This project includes a multi-disciplinary approach, utilizing X-ray crystallography, biochemistry, biophysics, and virology, in a collaborative effort among investigators from four different institutions. Research outcomes will augment the understanding of functional domain utilization for viral infection in general, particularly for non-enveloped viruses that must evolve strategies for traversing the complex cellular milieu en route to and from their sites of replication, and provide further insight into the mechanism(s) of viral adaptations to new hosts, a phenomenon that often gives rise to emerging viral pathogens.Broader Impacts: The research objectives of this project will be integrated into the educational training of high school, undergraduate and graduate students in the use of a multidisciplinary approach for studies aimed at deciphering structure-function correlations for macromolecular interactions. Specifically, high school students will be recruited to work on this project during the summer months through the University of Florida Student Science Training Program (SSTP), three undergraduate students are already actively involved in this project, and three of the specific research aims will form the research component of the Ph.D thesis of a graduate student. Undergraduate and graduate students will also participate in the research studies to be conducted in the collaborating laboratories. In addition, a specific outreach aim will involve the production of an educational CD-ROM titled "Structural biology as a tool for scientific research" as part of the University of Florida "Excursion in Science" Program web based learning tools aimed at K-12 students. This aim will also include the development of a three day workshop as part of the University of Florida "Teachers as Scholars" summer program to accompany the CD-ROM that will provide interdisciplinary physics, math, and biology high school teachers with hands-on experience and theoretical knowledge of structural biology that can be integrated into their classroom teaching strategies.

组装大分子的三维（3D）结构以实现功能性利用。 然而，这种3D结构一旦被顺应，就能够根据需要进行转变和/或重新排列，以实现必要的功能相互作用。 在宿主细胞感染过程中，定义病毒生命周期的连续大分子相互作用代表了大分子结构功能利用的高保真度的极好例子。 然而，尽管对病毒生命周期进行了大量研究，但关于促进衣壳-宿主相互作用、结构衣壳转换和衣壳-基因组相互作用的3D结构信息有限。 该项目将利用小鼠微小病毒（MVM）（ssDNA细小病毒科成员）的天然变异体和新出现的强毒株作为病毒模型，以破译成功的宿主细胞受体识别，细胞运输和基因组相互作用引发的衣壳成熟的结构要求。 该模型系统也是研究病毒株的理想选择，这些病毒株彼此相似，但由于其衣壳病毒蛋白的微小结构变异或氨基酸差异而在其宿主范围、组织嗜性和感染的致病性结果方面存在显著差异，这些差异改变了所需的感染性相互作用。 这些变异通常会改变病毒宿主-细胞相互作用，包括感染性受体利用，这在新出现的病毒中很常见。 该项目包括多学科的方法，利用X射线晶体学，生物化学，生物物理学和病毒学，在来自四个不同机构的研究人员之间的合作努力。 研究成果将增强对病毒感染的功能域利用的理解，特别是对于无包膜病毒，这些病毒必须进化出穿越复杂细胞环境的策略，以往返于其复制位点，并提供对病毒适应新宿主的机制的进一步了解，这种现象通常会导致新出现的病毒病原体。该项目的研究目标将被纳入高中，本科和研究生的教育培训中，使用多学科方法进行旨在破译大分子相互作用的结构-功能相关性的研究。 具体来说，高中生将被招募到这个项目在夏季的几个月里通过佛罗里达大学学生科学培训计划（SSTP），三个本科生已经积极参与这个项目，和三个具体的研究目标将形成博士论文的研究组成部分的研究生。 本科生和研究生也将参加在合作实验室进行的研究。 此外，一个具体的推广目标将涉及制作一个教育光盘，题为“结构生物学作为科学研究的工具”，作为佛罗里达大学“科学游览”计划的一部分，以K-12学生为目标的基于网络的学习工具。 这一目标还将包括一个为期三天的研讨会的发展，作为佛罗里达大学“教师作为学者”夏季计划的一部分，以配合光盘，将提供跨学科的物理，数学和生物学高中教师与实践经验和理论知识的结构生物学，可以整合到他们的课堂教学策略。