Inhibition of intracellular cAMP-dependent protein kinase using mutant genes of the regulatory type I subunit.
Inhibition of intracellular cAMP-dependent protein kinase using mutant genes of the regulatory type I subunit.
复制标题
DOI：
10.1016/s0021-9258(18)45175-7
复制
发表时间：
1987-09
期刊：
影响因子：
--
通讯作者：
C. Clegg;L. A. Correll;G. Cadd;G. McKnight
中科院分区：
文献类型：
--
作者：
C. Clegg;L. A. Correll;G. Cadd;G. McKnight
构建了编码环amp依赖性蛋白激酶调控型1亚基（RI）突变形式的表达载体。这些突变改变了一个特定的氨基酸，该氨基酸存在于RI蛋白的两个同源camp结合域中。当这些表达载体被引入NIH 3T3和Y1肾上腺皮质肿瘤细胞时，产生了一个突变的RI蛋白，该蛋白以显性方式起作用，导致camp依赖性蛋白激酶激活的20-400倍抑制。此外，肾上腺细胞中cAMP控制的过程被阻断；细胞对cAMP的生长抑制作用产生抗性，并且类固醇合成缺陷。突变体RI基因在细胞中的表达为探索cAMP和蛋白磷酸化在细胞内信号传导过程中的作用提供了一种特定的手段。
Expression vectors were constructed that code for mutated forms of the regulatory type 1 subunit (RI) of cyclic AMP-dependent protein kinase. These mutations alter a specific amino acid which is present in each of two homologous cAMP-binding domains of the RI protein. When these expression vectors were introduced into NIH 3T3 and Y1 adrenocortical tumor cells a mutant RI protein was produced that acted in a dominant fashion to cause a 20-400-fold inhibition of cAMP-dependent protein kinase activation. In addition, processes controlled by cAMP in adrenal cells were blocked; cells became resistant to the growth-inhibitory effects of cAMP and defective in steroid synthesis. Expression of mutant RI genes in cells provides a specific means to explore the role of cAMP and protein phosphorylation in the process of intracellular signalling.