Structural and Biochemical Studies of Crenarchael Viral Proteins

像SSV和STIV这样的新城疫病毒存在于极端高温(175至200华氏度)和酸性(1.0pH 3.0)环境中，比如在黄石国家公园发现的那些环境。虽然SSV和STIV是特征最好的新城疫病毒之一，但我们对这些独特的嗜热病毒的了解有限。例如，SSV和STIV基因组的序列分析无法确定大多数假定基因的同源基因。因此，大多数病毒蛋白还没有被赋予任何功能，这些蛋白在病毒生命周期中扮演的角色仍然是一个谜。该项目使用结构生物学和生物化学来(I)研究SSV和STIV蛋白质组的功能作用。这些蛋白质的结构将用X射线结晶学来确定。在许多情况下，结构与蛋白质数据库中的结构的相似性有望表明病毒蛋白质的功能。此外，该项目寻求(Ii)识别与特定病毒蛋白相互作用形成高阶复合体的病毒或宿主蛋白或DNA序列。鉴定这些相互作用的大分子将提供进一步的功能洞察。总之，这些努力将有助于在分子水平上更好地了解SSV和STIV的生命周期，因此该项目是朝着了解居住在这些极端环境中的独特病毒迈出的重要一步。此外，这些病毒非常令人感兴趣，因为它们是理解高温下生命所需的生物化学和分子生物学的模型系统。通过它们与宿主机器的相互作用，它们为研究重要的生化事件提供了入口点，从而打开了了解仍然鲜为人知的克伦考古群最基本过程的窗口。从历史上看，病毒也是生物技术的新型酶的来源，特别是参与核酸生物合成和修饰的酶。因此，鉴定新城疫病毒耐热酶可能在生物技术中有重要的应用。最后，从生命的三个领域的病毒的比较研究中获得的见解将有助于更好地了解病毒的起源和进化。广泛的影响：蒙大拿州立大学热生物学研究所(TBI)内正在进行的大部分工作正通过TBI推广计划向普通公众传播。关于嗜热病毒的工作是全球环境倡议的重点之一，因此也是这些外联工作的一个主要重点。外展的合作伙伴包括国家公园管理局/黄石国家公园，并涉及准备和审查供公园内公开展示的材料。公园系统和普通公众一样，越来越意识到居住在这些世界著名的热特征中的独特微生物，并正在努力增加公园内描述这些微生物的教育标志。这项研究与PI、系、大学和NSF的教育目标有很强的整合。高中、本科生、博士后和博士后将接受生物化学、结构生物学和热生物学方面的重要培训。鼓励代表人数不足的群体参与，特别是妇女和美洲土著人，他们的参与是有据可查的。

Crenarchaeal viruses such as SSV and STIV exist in extreme thermal (175 to 200 degrees Fahrenheit) and acidic (1.0 pH 3.0) environments such as those found in Yellowstone National Park. While SSV and STIV are among the best characterized Crenarchaeal viruses, our understanding of these unique hyperthermophilic viruses is limited. For example, sequence analysis of the SSV and STIV genomes fails to identify homologues for the majority of the putative genes. Hence, most viral proteins have yet to be assigned any function, and the roles that these proteins play in the viral life cycles remains a mystery. This project uses structural biology and biochemistry to (i) investigate the functional roles of the SSV and STIV proteomes. The structures of these proteins will be determined using X-ray crystallography. In many cases, the similarity of a structure to those in the Protein Data Bank is expected to suggest a function for the viral protein. In addition, this project seeks to (ii) identify viral or host proteins, or DNA sequences, that interact with specific viral proteins to form higher order complexes. Identification of these interacting macromolecules will provide further functional insight. Together, these efforts will contribute to an increased understanding of the SSV and STIV life cycles at the molecular level and thus this project represents a significant step toward understanding the unique viruses that inhabit these extreme environments. In addition, these viruses are of great interest because they serve as model systems for understanding the biochemistry and molecular biology required for life at high temperatures. Through their interactions with host machinery, they provide entry points for the study of important biochemical events, thus opening windows onto the most fundamental processes of the still poorly understood Crenarchaeota. Historically, viruses have also been a source of novel enzymes for biotechnology, particularly enzymes involved in nucleic acid biosynthesis and modification. Thus, identification of thermostable enzymes from the Crenarchaeal viruses might have important applications in biotechnology. Finally, the insights to be gained from comparative investigation of viruses from all three domains of life will be helpful in providing a better understanding of viral genesis and evolution.Broader Impacts: Much of the ongoing work within the Thermal Biology Institute (TBI) at Montana State University is being communicated to the general public by TBI outreach programs. Work on hyperthermophilic viruses is one of the focal points within TBI and is thus a major focus of these outreach efforts. Partners in outreach include the National Park Service /Yellowstone National Park, and involve preparation and review of materials for public presentation within the Park. The Park System is increasingly aware, as is the general public, of the unique microbial organisms inhabiting these world famous thermal features, and is working to increase the educational signage within the park that describes these organisms. There is strong integration of the research with the educational goals of the PI, the Department, the University, and the NSF. Students at the high school, undergraduate, doctoral and post-doctoral levels will receive significant training in biochemistry, structural biology and thermal biology. The participation of under represented groups, particularly women and Native Americans is encouraged, and their participation is well documented.