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

Vierling：这是power研究增强奖。本项目的直接目标是建立一种遗传学方法，利用蓝细菌Synechocystis sp. PCC 6803，对一类新的分子伴侣小热休克蛋白（sHSPs）进行结构和功能分析。这些普遍存在于真核生物和原核生物中的应激蛋白，在几乎所有生物的热应激过程中积累，并且似乎在许多生物的某些发育和生长阶段转变中也起作用。虽然存在sHSP的工作模型，但基本上没有关于sHSP作用机制细节的数据，而且很少有直接的体内数据来支持该模型。最近，京都大学的H. Fukuzawa博士发现，在藻胞菌HSP16.6中，单个sHSP基因的缺失会导致无法产生耐热性，但不会影响正常的细胞生长。这是首次证明原核生物的sHSP是高温耐受性所必需的，并且是除真菌神经孢子菌外的任何生物中第一个与sHSP突变相关的表型。由于聚胞菌可以很容易地从遗传学上操纵，这一发现为详细分析sHSP的结构和功能提供了极好的机会。胞囊菌的另一个优势是它的全基因组已经被测序。该项目将与日本京都大学的H. Fukuzawa合作进行。该奖项为Vierling博士实验室的蓝藻培养所需的设备购置提供了支持。购置这些设备对这个项目的启动至关重要。＊＊＊

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. ***