STRUCTURE OF HEAT SHOCK TRANSCRIPTION FACTOR FROM YEAST

本项目旨在研究热休克转录的结构， 因子（HSF）从酵母，特别是集中在形成的复合物 在HSF和它的DNA结合位点之间，热休克元件（HSE）。 HSF是 这是一个重要的自我平衡系统的一部分，称为热休克反应， 其用于保护细胞免受升高的 温度和某些其他环境压力。 在酵母中，HSF是一种 组成型和热诱导转录的必需蛋白 激活，其转录活性受后 翻译磷酸化 此外，HSF可能作为一种 三聚体，一种新的寡聚态的DNA结合蛋白，并可能 含有新的DNA结合基序。 因为， 热休克反应的转录调控已经高度 在所有真核生物中保守，彻底理解 酵母HSF的结构将适用于我们的理解 所有生物体都有热休克反应。 这项研究的长期目标之一将是研究X射线 晶体学研究了HSE和A之间形成的复合物的结构。 含有DNA结合区的HSF片段。 为此， 将使用生物物理、生物化学和分子生物学技术 分析HSE站点的顺序要求和域结构 和HSF的寡聚状态。 有了这些信息， 可以制备寡核苷酸和蛋白质片段用于结晶 审判 同时，对蛋白质决定簇的诱变研究 DNA分子识别所必需的一系列步骤将被启动。 这 研究将补充最终的晶体结构。

This project aims to study the structure of the heat shock transcription factor (HSF) from yeast, specifically focussing on the complex formed between HSF and its DNA binding site, the heat shock element (HSE). HSF is part of an important homeostatic system called the heat shock response, which serves to protect the cells from damage caused be elevated temperatures and certain other environmental stresses. In yeast, HSF is an essential protein for both constitutive and heat-inducible transcriptional activation, and its transcriptional activity is regulated by post- translational phosphorylations. Furthermore, HSF probably exists as a trimer, a novel oligomeric state for a DNA binding protein, and might contain a new DNA binding motif. Because the basic mechanism of transcriptional regulation of the heat shock response has been highly conserved throughout all eukaryotes, a thorough comprehension of the structure of the yeast HSF will be applicable to our understanding of the heat shock response in all organisms. One of the long term objectives of this research will be to study X-ray crystallography the structure of the complex formed between HSE and a fragment of HSF containing the DNA binding region. To this end, biophysical, biochemical and molecular biological techniques will be used to analyze the sequence requirements for HSE sites and the domain structure and oligomeric state of HSF. With this information, the appropriate oligonucleotide and protein fragments can be prepared for crystallization trials. concurrently, a mutagenic study of the protein determinants necessary for the molecular recognition of the DNA will be initiated. This study will complement the eventual crystal structure.