REGULATION OF ENDOTHELIN-1 GENE EXPRESSION

内皮素-1（ET-1）是一种由21个氨基酸组成的多肽， 内皮细胞 它既是一种有效的血管收缩剂， 因子 内皮素-1在血管生理中的作用及其参与 血管疾病不详。 拟议工作的目的包括使用 基因研究，以更好地了解ET-1的功能，使用 该基因作为研究内皮细胞的模型系统 ET-1调节序列在内皮细胞转录中的应用 转基因动物中的细胞靶向。 我们将阐明ET-1基因的顺式作用DNA元件， 通过转染与报告基因连接的ET-1序列体外表达。 这些 实验将通过体内类似研究进行补充， 携带ET-1-lacZ转基因的转基因小鼠。 体外足迹法 和凝胶位移分析将允许表征反式作用 结合这些序列的蛋白质。 ET-1顺式作用序列， 将采用似乎对内皮细胞转录特异的 DNA结合蛋白的纯化和克隆。 要素 ET-1启动子，其提供高水平的内皮细胞表达， 转基因动物将用于开发转基因小鼠品系 其表达血液中ET-1水平增加或减弱 血管壁 这些小鼠将被评估血液中的变化 压力和血管疾病的易感性。 内皮细胞特异性克隆的可用性 反式作用因子将使研究 这一重要细胞类型的发育和表型多样性。 的 指导内皮细胞表达的构建体的可用性 转基因小鼠将允许一种方法来产生小鼠模型， 血管疾病

Endothelin-1 (ET-1) is a 21-amino acid peptide synthesized in endothelial cells. It is both a potent vasoconstrictor and growth factor. The role of ET-1 in vascular physiology and its involvement in vascular disease is unknown. Aims of the proposed work include the use of genetic studies to better understand the function of ET-1, the use of this gene as a model system for the study of endothelial cell transcription, and the use of ET-1 regulatory sequences for endothelial cell targeting in transgenic animals. We will elucidate the cis-acting DNA elements of the ET-1 gene in vitro by transfection of ET-1 sequences linked to a reporter gene. These experiments will be complemented by analogous studies in vivo using transgenic mice which carry ET-1-lacZ transgenes. In vitro footprinting and gelshift analysis will allow characterization of trans-acting proteins which bind these sequences. ET-1 cis-acting sequences which appear to be specific for endothelial cell transcription will be employed in the purification and cloning of DNA binding proteins. Elements of the ET-1 promoter which provide high level endothelial cell expression in transgenic animals will be utilized to develop lines of transgenic mice which express either increased or attenuated levels of ET-1 in the blood vessel wall. These mice will be evaluated for alterations in blood pressure and predisposition for vascular disease. The availability of a clone for an endothelial cell-specific trans-acting factor will make possible studies investigating the development and phenotypic diversity of this important cell type. The availability of constructs which direct endothelial cell expression in transgenic mice will allow an approach to the generation of mouse models of vascular disease.