NGF-INDUCED DEREPRESSION OF PERIPHERIN GENE EXPRESSION

我的研究兴趣是组织特异性转录调控 基因表达和细胞向特定细胞定向的机制 分化途径 PC 12细胞神经元分化的诱导 神经生长因子（NGF）是研究这些细胞的模型系统。 问题. 为了了解神经生长因子触发神经细胞凋亡的机制， 神经元分化，我们正在研究外周蛋白的调节 基因，一种神经特异性中间丝蛋白， 在神经元分化时转录诱导。 以前我们已经定义了近端;和远端的积极因素， 外周蛋白启动子，其是由NGF诱导所必需的，以及 负调控元件（NRE），其在抑制 在未分化和非神经元细胞中的外周蛋白活性。 我们 目前的重点是确定积极和消极的转录因子 与这些调控区相互作用。 该提案将重点关注 外周蛋白的负调节通过阻遏蛋白结合到 NRE。 一个主要的目标是纯化和克隆阻遏蛋白， 确定神经生长因子介导的机制如何改变其表达和活性。 此外，拥有克隆的阻遏蛋白将有助于 使用体外转录来确定其作用水平的研究，以及 作为鉴定蛋白质阻遏物结构域的结构研究。 最后，研究了NRE突变对一种表达模式的影响。 外周蛋白启动子-lacZ融合基因在转基因小鼠中的表达将表明 NRE对神经元特异性表达的体内重要性。

My research interests are the transcriptional regulation of tissue-specific gene expression and the mechanism of commitment of a cell to a particular differentiation pathway. The induction of neuronal differentiation in PC12 cells by nerve growth factor (NGF) is a model system for studying these questions. In order to understand the mechanism by which NGF triggers neuronal differentiation, we are studying the regulation of the peripherin gene, a neural-specific intermediate filament protein which is transcriptionally induced at the time of neuronal differentiation. Previously we have defined proximal; and distal positive elements in the peripherin promoter which are necessary for induction by NGF, as well as a negative regulatory element (NRE) which has a functional role in repressing peripherin activity in undifferentiated and non-neuronal cells. Our present focus is to identify positive and negative transcription factors which interact with these regulatory regions. This proposal will focus on the negative regulation of peripherin by repressor proteins binding to the NRE. A major goal is to purify and clone the repressor protein in order to determine how NGF-mediated mechanisms modify its expression and activity. In addition, possession of the cloned repressor protein will facilitate studies using in vitro transcription to define its level of action, as well as structural studies identifying the repressor domain of the protein. Finally, the effect of mutation at the NRE on the expression pattern of a peripherin promoter-lacZ fusion gene in transgenic mice will indicate the in vivo importance of the NRE for neuronal-specific expression.