EUKARYOTIC CHROMATIN STRUCTURE AND GENE REGULATION

本课组持续开展染色质结构和热休克基因表达调控的研究，重点关注热休克转录因子（HSF）的结构和功能以及热休克基因启动子在染色质中的重塑。酵母双杂交筛选已经确定了果蝇HSF的几个相互作用蛋白。目前正在对这些蛋白进行表征，并通过分子和细胞分析测试其对HSF可能的调节作用。atp依赖性核小体重塑因子（NURF）最大亚基的克隆已经完成。该亚基被发现由一个非常大的ORF编码，预测一个新的301 kD多肽，包含一个溴结构域和三个PHD指。我们最近成功地重建了301K ORF的全长cDNA。目前，我们正在表征这种新蛋白，并试图通过杆状病毒表达载体的共表达，从所有四个亚基重建NURF活性，以确定NURF核小体重塑活性所需的最小组分。-染色体，果蝇，基因，热休克，核小体，转录，

Our group has continued studies of chromatin structure and the regulation of heat shock gene expression, with emphasis on the structure and function of the heat shock transcription factor (HSF) and on the remodeling of the heat shock gene promoter in chromatin. Yeast two-hybrid screens have identified several interacting proteins for Drosophila HSF. These proteins are currently being characterized and tested for possible regulatory effects on HSF by molecular and cellular assays. The cloning of the largest subunit for the ATP-dependent nucleosome remodeling factor (NURF) has been completed. This subunit was found to be encoded by a very large ORF predicting a novel, 301 kD polypeptide containing a bromodomain and three PHD fingers. We recently succeeeded in reconstructing a full length cDNA for the 301K ORF. Currently we are characterizing this novel protein and attempting to reconstitute NURF activity from all four subunits by co-expression from baculovirus expression vectors to determine the minimal components necessary for the nucleosome remodeling activity of NURF. - Chromosome, Drosophila, Gene, Heat Shock, Nucleosome, Transcription,