NGF-INDUCED DEREPRESSION OF PERIPHERIN GENE EXPRESSION

我的研究兴趣是组织特异性转录调控 基因表达与细胞对特定物质的承诺机制 分化途径。PC12诱导神经分化的实验研究 神经生长因子(NGF)诱导的细胞是研究这些问题的一个模型系统。 问题。为了了解NGF触发的机制 神经元分化，我们正在研究外周蛋白的调节 基因，一种神经特异的中间丝蛋白，它是 在神经元分化时被转录诱导。 在此之前，我们已经定义了近端和远端的正性元素 外周蛋白启动子是NGF诱导所必需的，以及一种 负性调节元件(NRE)具有抑制作用 未分化和非神经细胞中的外周蛋白活性。我们的 目前的重点是识别正转录因子和负转录因子 它们与这些监管区域相互作用。这项提案将重点放在 抑制蛋白与外周蛋白结合对外周蛋白的负性调节 没有。一个主要的目标是纯化和克隆阻遏蛋白，以便 确定NGF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也是如此 因为结构研究确定了蛋白质的阻遏结构域。 最后，NRE突变对A基因表达模式的影响 外周蛋白启动子-LacZ融合基因在转基因小鼠中的表达 NRE在体内对神经元特异性表达的重要性。

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.