CAREER: Structural and Biochemical Studies of Essential Viral Proteins

WuttkeMCB 98756631。本研究的目的是利用结构和生物化学方法研究3AB蛋白及其裂解产物、3A和3B蛋白这三种病毒必需蛋白的结构和功能之间的关系。脊髓灰质炎病毒3AB蛋白是一种重要的多功能蛋白，在病毒复制中发挥着几个关键作用。多维异核核磁共振波谱将用于确定人类脊髓灰质炎病毒3A和3AB蛋白的高分辨率溶液结构。3AB与其生物伙伴病毒聚合酶之间相互作用的基本化学本质将使用生化、生物物理和结构方法进行探索。最后，将使用磁场中生物分子排列的新方法获得的偶极耦合来精炼3A和3AB结构。由于3A和3AB蛋白存在于所有微小核糖核酸病毒中，对它们的结构和功能的深入了解将有助于我们深入了解许多重要病毒系统的基本生物化学，并促进我们对蛋白质/蛋白质相互作用如何调节生物活性的理解。该教育项目的目标是将生物信息学和生物分子结构分析纳入科罗拉多大学博尔德分校的本科生物化学课程。生物信息学工具的使用，如序列比较、全基因组序列分析以及生物分子结构的可视化和操纵，是现代生物化学培训的基本组成部分。互联网上可用的信息和工具的爆炸性增长，以及过去几年出现的三维结构的数量表明，随着时间的推移，生物化学的这一方面的影响只会越来越大。该项目将开发必要的工具，将这一强大的生物化学新领域纳入到生物化学本科生和研究生的培训中，以便为他们在专业学校和行业的未来做好准备。2一般概述该研究计划的目标是使用结构和生物化学方法来研究三种基本病毒蛋白，3AB蛋白及其裂解的后代，3A和3B蛋白的结构和功能之间的关系。这些蛋白质是由一个重要的病毒家族--小核糖核酸病毒产生的，其成员包括脊髓灰质炎和普通感冒的病原体。这些蛋白质的三维结构将与它们的生物学功能有关。将使用生化、生物物理和结构方法来探索这些蛋白质与它们相互作用的蛋白质伙伴之间相互作用的基本化学性质。由于3A和3AB蛋白存在于所有微小核糖核酸病毒中，对它们的结构和功能的深入了解将有助于我们深入了解许多重要病毒系统的基本生物化学，并促进我们对蛋白质/蛋白质相互作用如何调节生物活性的理解。该教育项目的目标是将生物信息学和生物分子结构分析纳入科罗拉多大学博尔德分校的本科生物化学课程。生物信息学工具的使用，如序列比较、全基因组序列分析以及生物分子结构的可视化和操纵，是现代生物化学培训的基本组成部分。互联网上可用的信息和工具的爆炸性增长，以及过去几年出现的三维结构的数量表明，随着时间的推移，生物化学的这一方面的影响只会越来越大。该项目将开发必要的工具，将这一强大的生物化学新领域纳入生物化学本科生和研究生的培训，以便为他们在专业学校和行业的未来做好准备。

WuttkeMCB 98756631. TECHNICAL ABSTRACTThe goals of the research program are to use structural and biochemical methods to investigate the relationship between structure and function of three essential viral proteins, the 3AB protein and its cleaved progeny, the 3A and 3B proteins. The poliovirus 3AB protein is an important, multifunctional protein that plays several critical roles in viral replication in members of the Picornaviridae family. Multidimensional heteronuclear NMR spectroscopy will be used to determine the high-resolution solution structure of the 3A and 3AB proteins from human poliovirus. The fundamental chemical nature of the interactions between 3AB and its biological partner, the viral polymerase, will be probed using biochemical, biophysical and structural methods. Finally, the 3A and 3AB structures will be refined using dipolar couplings obtained by new methods for biomolecular alignment in a magnetic field. Because the 3A and 3AB proteins are found in all picornaviruses, a better understanding of their structures and functions will provide important insights into the fundamental biochemistry of a large number of important viral systems, as well as advance our understanding of how protein/protein interactions modulate biological activity. The aims of the education program are to integrate bioinformatics and biomolecular structure analysis into the undergraduate biochemistry curriculum at the University of Colorado at Boulder. The use of bioinformatics tools, such as sequence comparison, analysis of complete genome sequences, and visualization and manipulation of biomolecular structures are essential components of training in modern biochemistry. The explosion of information and tools available over the Internet, and the number of three-dimensional structures that have become available in the last few years indicate this aspect of biochemistry is only going to increase in impact over time. This project will develop the tools needed to incorporate this powerful new area of biochemistry into the training of biochemistry undergraduate and graduate students in order to prepare them for their future in professional schools and industry.2 GENERAL ABSTRACTThe goals of the research program are to use structural and biochemical methods to investigate the relationship between structure and function of three essential viral proteins, the 3AB protein and its cleaved progeny, the 3A and 3B proteins. These proteins are produced by an important family of viruses, the picornaviruses, whose members include the causative agents of polio and the common cold. The three-dimensional structure of these proteins will be related to their biological functions. The fundamental chemical nature of the interactions between these proteins and the protein partners they interact with will be probed using biochemical, biophysical and structural methods. Because the 3A and 3AB proteins are found in all picornaviruses, a better understanding of their structures and functions will provide important insights into the fundamental biochemistry of a large number of important viral systems, as well as advance our understanding of how protein/protein interactions modulate biological activity. The aims of the education program are to integrate bioinformatics and biomolecular structure analysis into the undergraduate biochemistry curriculum at the University of Colorado at Boulder. The use of bioinformatics tools, such as sequence comparison, analysis of complete genome sequences, and visualization and manipulation of biomolecular structures are essential components of training in modern biochemistry. The explosion of information and tools available over the Internet, and the number of three-dimensional structures that have become available in the last few years indicate this aspect of biochemistry is only going to increase in impact over time. This project will develop the tools needed to incorporate this powerful new area of biochemistry into the training of biochemistry undergraduate and graduate students in order to prepare themfor their future in professional schools and industry.