HEAT SHOCK PROTEIN 27 EXPRESSION IN OSTEOBLASTS

这一应用将定义热休克蛋白的分子基础 (HSP)27在成骨细胞中的表达，基于以下假设 成骨细胞中热休克蛋白27的结构性表达可能是一个主要的决定因素 由于成骨细胞具有相对的耐热性，因此对压力的耐受性较差。 具体目标1旨在确定多个热休克蛋白的来源 27个在应激和非应激状态下表达的相关蛋白质和转录本 MC3T3细胞中不同分化状态的成骨细胞基因组学 相关成绩单的来源将由传统的 筛选和序列分析。《特定目标2》将调查 结构型HSP 27表达的机制，热应激和 雌激素对热休克蛋白27表达的调控模式。我们将进行比较 在应激和非应激细胞之间。转录调控将 将在报告基因结构中进行检查并进行比较 在MCTC929成纤维细胞之间，阴性对照细胞系不 表达了HSP27，并在小鼠mC3T3细胞系中进行了研究 经历了节目的差异化。《特定目标3》将考察潜力 热休克蛋白27与其他细胞相互作用的研究 蛋白质，基于热休克蛋白分子作为分子的已知功能 监护人。此处未知蛋白质(由非应激或热的cdna编码) 与外体相互作用的成骨细胞)表达HSP27 酵母菌将通过核质相互作用选择策略进行鉴定 克隆技术。

This application will define the molecular basis of heat shock protein (hsp) 27 expression in osteoblasts, based on the hypothesis that the constitutive expression of HSP 27 in osteoblasts may be a major determinant of stress tolerance as the osteoblast is relatively thermal resistant. Specific aim 1 is directed towards identifying the origins of multiple HSP 27 related proteins and transcripts expressed in stressed and unstressed osteoblasts of various differentiated states in MC3T3 cells. Genomic origins of the related transcripts will be defined by conventional screening and sequence analysis. Specific aim 2 will investigate the mechanisms underlying constitutive hsp 27 expression, heat stressed and estrogen regulated modes of hsp 27 expression. Comparisons will be made between stressed and non stressed cells. Transcriptional regulation will be examined in reporter gene constructs and comparisons will be made between MCTC929 fibroblasts, a negative control cell line that does not express hsp 27, and studies carried out in the mouse mC3T3 cell line that undergoes program differentiation. Specific aim 3 will examine potential functions of hsp 27 by determining hsp 27 interactions with other cellular proteins, based upon known functions of hsp molecules as molecular chaperones. Here unknown proteins (encoded by cDNA for nonstressed or heat stressed osteoblasts) which interact with episomal expressed hsp 27 in yeast will be identified by a karyoplasmic interaction selection strategy of cloning.