ISOLATION AND USES OF CGMP PHOSPHODIESTERASE CDNAS

环核苷酸磷酸二酯酶是细胞周期中的关键成分。 CAMP和cGMP介导的信号转导通路的调控。 这种酶家族的大小及其调控的复杂性 可能得不到充分的赏识。研究它们的重要性的根源 从它们内在的生化和生物兴趣以及从 他们是抗哮喘，抗血栓， 强心剂和抗抑郁药。尽管已经在 多年来的生化水平、某些异构体的cDNA和基因 已经很难克隆了，这现在已经成为一种必需品。 我们已经开发出一种基因选择程序，可以恢复 编码这些酶的DNA。已经创造了这样的条件 能表达功能蛋白的大肠杆菌Lambda溶原菌 环核苷酸或cGMP磷酸二酯酶可以生长。这种方法已经被 用于恢复可能编码cGMP刺激的cGMP特异的cDNA 盘基网眼菌的磷酸二酯酶。CGMP介导的信号转导 转导级联和这种磷酸二酯酶调节细胞的运动 并将被用来询问cGMP是如何介导 信号转导控制细胞骨架的动员和其他 具有发展意义的重大事件。相同的遴选程序将是 改进和扩展以恢复选定的哺乳动物磷酸二酯酶cDNA 并对其分子、生化和药理性质进行研究。

The cyclic nucleotide phosphodiesterases are crucial components in the regulation of the signal transduction cascades mediated by cAMP and cGMP. The size of this family of enzymes and the complexity of their regulation may not be adequately appreciated. The importance of studying them stems from their intrinsic biochemical and biological interest and from the fact that they are the targets of anti-asthmatic, anti-thrombolytic, cardiotonic, and anti-depressant drugs. Although they have been studied at the biochemical level for many years, the cDNAs and genes of some isoforms have been difficult to clone and this has now become a necessity. We have developed a genetic selection procedure that allows recovery of cDNAs coding for these enzymes. Conditions have been created in which lambda lysogens of E. coli that are capable of expressing a functional cyclic nucleotide or cGMP phosphodiesterase can grow. This method has been used to recover cDNAs that may code for a cGMP stimulated cGMP specific phosphodiesterase from Dictyostelium discoideum. The cGMP mediated signal transduction cascade and this phosphodiesterase regulate the motility of this organism and will be used to ask questions about how cGMP mediated signal transduction controls mobilization of the cytoskeleton and other developmentally important events. The same selection procedure will be improved and extended to recover selected mammalian phosphodiesterase cDNAs and to study their molecular, biochemical, and pharmacological properties.