Abraham I, Hunter R J, Sampson K E, Smith S, Gottesman M M, Mayo J K
Cell Biology Department, Upjohn Co., Kalamazoo, Michigan 49001.
Mol Cell Biol. 1987 Sep;7(9):3098-106. doi: 10.1128/mcb.7.9.3098-3106.1987.
The isolation of mutant cell lines affecting the activity of cyclic AMP (cAMP)-dependent protein kinase (PK-A) has made it possible to determine the function of this kinase in mammalian cells. We found that both a CHO cell mutant with a defective regulatory subunit (RI) for PK-A and a transfectant cell line expressing the same mutant kinase were sensitive to multiple drugs, including puromycin, adriamycin, actinomycin D, and some antimitotic drugs. The mutant and transfectant cells, after treatment with a concentration of the antimitotic drug colcemid that had no marked effect on the wild-type parent cell, had a severely disrupted microtubule network. The phenotype of hypersensitivity to the antimitotic drug colcemid was used to select revertants of the transfectant and the original mutant. These revertants simultaneously regained normal multiple drug resistance and cAMP sensitivity, thus establishing that the characteristics of colcemid sensitivity and cAMP resistance are linked. Four revertants of the transfectant reverted because of loss or rearrangement of the transfected mutant RI gene. These revertants, as well as one revertant selected from the original mutant, had PK-A activities equal to or higher than that of the parent. In these genetic studies, in which linkage of expression of a PK-A mutation with drug sensitivity is demonstrated, it was established that the PK-A system is involved in regulating resistance of mammalian cells to multiple drugs.
影响环磷酸腺苷(cAMP)依赖性蛋白激酶(PK-A)活性的突变细胞系的分离,使得确定该激酶在哺乳动物细胞中的功能成为可能。我们发现,一种具有缺陷的PK-A调节亚基(RI)的CHO细胞突变体和一种表达相同突变激酶的转染细胞系,对多种药物敏感,包括嘌呤霉素、阿霉素、放线菌素D和一些抗有丝分裂药物。在用对野生型亲本细胞无明显影响的抗有丝分裂药物秋水仙酰胺处理后,突变体和转染细胞的微管网络严重受损。利用对秋水仙酰胺这种抗有丝分裂药物的超敏表型来筛选转染细胞和原始突变体的回复突变体。这些回复突变体同时恢复了正常的多重耐药性和cAMP敏感性,从而证实秋水仙酰胺敏感性和cAMP耐药性的特征是相关联的。转染细胞的四个回复突变体是由于转染的突变RI基因的缺失或重排而回复的。这些回复突变体,以及从原始突变体中筛选出的一个回复突变体,其PK-A活性等于或高于亲本。在这些证明PK-A突变表达与药物敏感性相关联的遗传学研究中,证实了PK-A系统参与调节哺乳动物细胞对多种药物的耐药性。