STRUCTURE OF HEAT SHOCK TRANSCRIPTION FACTOR FROM YEAST

本项目旨在研究热休克转录的结构。 来自酵母的因子(HSF)，专门关注形成的复合体 HSF与其DNA结合部位之间的热休克元件(HSE)。HSF是 这是一个重要的平衡系统的一部分，称为热休克反应， 它起到保护细胞免受损伤的作用 温度和某些其他环境压力。在酵母中，HSF是一种 构成性和热诱导转录所必需的蛋白质 激活，其转录活性受转录后蛋白调节。 翻译磷酸化。此外，HSF可能以 三聚体，DNA结合蛋白的一种新的寡聚体状态，并可能 包含一个新的DNA结合基序。因为它的基本机制是 热休克反应的转录调控一直很高 在所有真核生物中都是保守的，对 酵母HSF的结构将适用于我们对 所有生物体的热休克反应。 这项研究的长期目标之一将是研究X射线 结晶学：HSE与a形成的络合物的结构 含有DNA结合区的HSF片段。为此， 将使用生物物理、生化和分子生物学技术 分析HSE站点和域结构的序列要求 和HSF的低聚状态。有了这些信息，适当的 可以制备用于结晶的寡核苷酸和蛋白质片段 审判。同时，对蛋白质决定因素进行了突变研究 将启动DNA分子识别所必需的。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.