Expression of Genes for Metabolism of Cyclic Adenosine 3′:5′-Monophosphate in Somatic Cells

作者：Alfred G. Gilman, John Dorrance Minna · 发表于：Journal of Biological Chemistry · 年份：1973 · DOI：10.1016/s0021-9258(19)43397-8 · 被引用次数：49 · 研究领域：CRISPR and Genetic Engineering、RNA Interference and Gene Delivery、Neuroscience and Neural Engineering

Abstract The effects of catecholamines (norepinephrine and isoproterenol) on the intracellular accumulation of cyclic adenosine 3':5'-monophosphate (cyclic AMP) were determined for clonal cell lines cultured in vitro. Catecholamine responsive (β+) and unresponsive (β-) clones were found. β+ responses were inhibited selectively by propranolol (1 µm) but not by practolol (10 µm), indicating that the response should be classified as β2+. These cell lines then were used as parents in somatic cell hybridization to study the genetic control of the β2+ response. In general, β2+ x β2+ matings yielded β2+ hybrid cells, while β2+ x β- matings yielded β- hybrid cells (less than 2% of the β2+ parental response). However, β2+ parental cells with large responses (300-fold stimulation) to catecholamine, when fused to β2+ parents with smaller (15-fold stimulation) responses, yielded hybrids with definite but minimal (average of 3-fold) stimulation. Chromosome analysis of all hybrid cells revealed no evidence for preferential loss of marker chromosomes of the β2+ parent. These data represent evidence for a heritable negative control mechanism regulating catecholamine responsiveness. In addition, the hybrid cell lines represent new biologic material for biochemical study of the macromolecular events between β-adrenergic stimulation and the accumulation of intracellular cyclic AMP.