Chinese Hamster Ovary Cells Transfected with the ras Oncogene or a DNA Sequence from a cAMP-Dependent Protein Kinase Mutant

Abstract

Ras oncogene products share similarities with the G proteins composing the adenylate cyclase system, which regulates cAMP levels within a cell. Chinese hamster ovary (CHO) cells with mutant cAMP-dependent protein kinase (with the nomenclature 10215 and 10248) as well as malignantly transformed and nontransformed wild type CHO cells (10001 and 10900, respectively) were transfected with the ras oncogene to determine if an alteration in the adenylate cyclase system and consequently in intracellular cAMP levels occurred. From subjective analysis it did not appear that this oncogene was able to alter cAMP sensitivity. This suggests that the intracellular cAMP levels within this mammalian cell system did not change. These results should only be taken as a preliminary step in analysis of the effects of the ras oncogene products. CHO cell lines 10001 and 10900 were also transfected with cosmids containing DNA from the cAMP-dependent protein kinase mutant 10215. This mutant as well as 10248 are not effected by elevated levels of cAMP. The transfection of 10001 and 10900 was performed to determine if cAMP resistant properties could be conferred to these cell lines. Only one of the two cosmids, cosmid 5, indicated any evidence of conveying cAMP resistance. Conclusive identification of cAMP properties with this cosmid is not yet definite, however.