HEAT SHOCK PROTEIN 27 EXPRESSION IN OSTEOBLASTS

这一应用将确定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HSP 27的组成性表达可能是一个主要的决定因素 因为成骨细胞相对耐热。 具体目标1是针对确定多种HSP的起源 27种相关蛋白和转录物在应激和非应激中表达 MC3T3细胞中各种分化状态的成骨细胞。 基因组 相关转录本的来源将由传统的 筛选和序列分析。 具体目标2将调查 组成性热休克蛋白27表达、热应激和 热休克蛋白27表达的雌激素调节模式。 将进行比较 在应激和非应激细胞之间。 转录调控将 将在报告基因构建体中进行检查并进行比较 MCTC 929成纤维细胞，阴性对照细胞系， 表达热休克蛋白27，以及在小鼠mC3T3细胞系中进行的研究， 进行程序分化。 具体目标3将审查 通过确定热休克蛋白27与其他细胞的相互作用 基于HSP分子作为分子的已知功能， 监护人 这里未知的蛋白质（由非应激或热应激的cDNA编码） 应激的成骨细胞）与表达HSP27的附加体相互作用， 酵母将通过核质相互作用选择策略鉴定 克隆。

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.