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.