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

柴胡病毒(Sulfolobus shibatae Virus，SSV)是一种新城疫病毒，存在于极端高温(大于80oC)和酸性(pH 3.0)的环境中，如在黄石国家公园发现的环境。虽然SSV1是新城疫病毒最好的特征，但目前对这些极其独特的病毒的了解非常有限。来自极端高温环境的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架(ORF)的同源物。因此，大多数SSV1病毒蛋白质组，34个开放阅读框中的30个，还没有被赋予任何功能，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射线结晶学，这个项目试图确定SSV1蛋白质组成员的结构同源物，从而提供对它们可能功能的洞察。这些结构研究的补充是为每一种蛋白质生产多克隆抗血清，并随后分析它们在病毒生命周期中的表达模式和功能。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：极端嗜热菌病毒可以作为模型系统，了解高温下生命所需的生物化学和分子生物学，因此，这项研究将有助于打开一扇窗，了解仍然鲜为人知的Crenarocota的一些最基本的过程。本项目所研究的结构生物学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出色的培训机会，包括为来自蒙大拿州的美洲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.