Structural Studies on the SSV1 Proteome; an Archeal Virus from an Extreme Thermal Environment

柴叶磺胺酸病毒（SSV）是一种绿古菌病毒，存在于极端高温（超过80摄氏度）和酸性（pH值3.0）环境中，例如黄石国家公园。虽然SSV1是所有绿古菌病毒中最具特征的，但目前对这些极其独特的病毒的了解非常有限。对来自极端热环境的ssv和大多数其他病毒的基因组序列分析未能在公开可用的数据库中找到绝大多数假定的开放阅读框（orf）的同源物。因此，大多数SSV1病毒蛋白质组，34个orf中的30个，尚未被赋予任何功能，这些蛋白质在病毒生命周期中扮演的角色仍然是一个谜。利用x射线晶体学，该项目旨在鉴定SSV1蛋白质组成员的结构同源物，从而深入了解其可能的功能。结构研究正在通过生产这些蛋白质的多克隆抗血清来补充，随后分析它们的表达模式和与病毒生命周期相关的功能。总之，这些数据将开始在分子水平上提供对SSV1生命周期的理解。更广泛的影响：极端嗜热细菌的病毒可以作为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，因此这项研究将有助于打开一扇窗口，了解一些仍然知之甚少的绿原菌的最基本的过程。本课题研究的结构生物学内容将纳入本科教学活动。此外，该项目将为不同层次的学生提供优秀的培训机会，包括为来自蒙大拿州的美国原住民学生提供暑期研究经验。

Sulfolobus shibatae Virus (SSV) is a Crenarchaeal virus that exists in extreme thermal (greater than 80oC) and acidic (pH 3.0) environments such as the ones found in Yellowstone National Park. While SSV1 is the best characterized of any Crenarchaeal virus, the current understanding of these extremely unique viruses is very limited. Genomic sequence analysis of SSVs and most other viruses from extreme thermal environments fails to find homologues in the publicly available data bases for the vast majority of the putative open reading frames (ORFs). Thus, most of the SSV1 viral proteome, 30 of 34 ORFs, have yet to be assigned any function, and the roles that these proteins play in the viral life cycle remains a mystery. Using X-ray crystallography, this project seeks to identify structural homologues for member of the SSV1 proteome, thus providing insight into their possible function. The structural studies are being supplemented by the production of polyclonal antisera for each of these proteins, with subsequent analysis of their expression patterns and function with respect to the viral life cycle. Together, these data will begin to provide an understanding of the SSV1 life cycle at the molecular level. Broader Impact: Viruses of extreme thermophiles can serve as model systems for understanding the biochemistry and molecular biology required for life at high temperatures, thus this research will help to open a window onto some of the most fundamental processes of the still poorly understood Crenarchaeota. The structural biology investigated in this project will be incorporated in the undergraduate educational activities. Additionally, this project will provide outstanding training opportunities to students at different levels, including summer research experience for Native American students from Montana.