POWRE: A Genetic Approach to Structure and Function Analysis of a New Class of Molecular Chaperones

9752978 Vierling这是POWRE研究增强奖。这个项目的近期目标是建立一种遗传方法，使用蓝藻聚胞藻。PCC 6803，用于分析一类新的分子伴侣--小分子热休克蛋白(SHSPs)的结构和功能。这些普遍存在的应激蛋白存在于真核生物和原核生物中，在几乎所有生物的热应激过程中积累，似乎也在许多生物的某些发育和生长阶段转换中发挥作用。虽然sHSP的工作模型已经存在，但基本上没有关于SHSP作用的机制细节的数据，也很少有直接的活体数据来支持该模型。最近，京都大学的H.Fukuzawa博士发现，蓝藻聚球藻HSP16.6中单个SHSP基因的缺失会导致无法形成耐热性，但不会影响正常细胞的生长。这是第一次证明原核生物的SHSP是耐高温所必需的，也是除真菌脉孢子菌以外的任何生物中第一个与SHSP突变相关的表型。由于聚孢子虫可以很容易地在基因上操纵，这一发现为详细分析SHSP的结构和功能提供了一个极好的机会。聚球藻的进一步优势是它的完整基因组已经测序。该项目将与日本京都大学的H.Fukuzawa合作进行。这项奖励为获得在Vierling博士的实验室培养蓝藻所需的设备提供了支持。获得这一设备对启动这一项目至关重要。***

9752978 Vierling This is a POWRE Research Enhancement Award. The immediate goal of this project is to establish a genetic approach, using the cyanobacterium Synechocystis sp. PCC 6803, for structure and function analysis of a new class of molecular chaperone, the small heat shock proteins (sHSPs). These ubiquitous stress proteins, found in both eukaryotes and prokaryotes, accumulate during heat stress in virtually all organisms, and appear to also have a role in certain developmental and growth stage transitions in many organisms. While a working model for the sHSPs exists, there are essentially no data on the mechanistic details of sHSP action, and very little direct in vivo data to support the model. Recently, Dr. H. Fukuzawa at Kyoto University discovered that deletion of the single sHSP gene in the cyanobacterium Synechocystis, HSP16.6, leads to inability to develop thermotolerance but does not affect normal cell growth. This is the first demonstration that a prokaryotic sHSP is required for high temperature tolerance, and is the first phenotype associated with an sHSP mutant in any organism other than the fungus Neurospora. Since Synechocystis can be easily manipulated genetically, this discovery affords an excellent opportunity for detailed analysis of sHSP structure and function. Synechocystis carries the further advantage that its complete genome has been sequenced. The project will be carried out in collaboration with H. Fukuzawa at Kyoto University in Japan. This award provides support toward the acquisition of equipment necessary for the culture of the cyanobacteria in Dr. Vierling's laboratory. The acquisition of this equipment is critical to the initiation of this project. ***