Ouabain Resistance Conferred by Expression of the cDNA for a Murine Na + , K + -ATPase α Subunit

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Abstract
The molecular basis for the marked difference between primate and rodent cells in sensitivity to the cardiac glycoside ouabain has been established by genetic techniques. A complementary DNA encoding the entire α 1 subunit of the mouse Na + - and K + -dependent adenosine triphosphatase (ATPase) was inserted into the expression vector pSV2. This engineered DNA molecule confers resistance against 10 -4 M ouabain to monkey CV-1 cells. Deletion of sequences encoding the carboxyl terminus of the α 1 subunit abolish the activity of the complementary DNA. The ability to assay the biological activity of this ATPase in a transfection protocol permits the application of molecular genetic techniques to the analysis of structure-function relationships for the enzyme that establishes the internal Na + /K + environment of most animal cells. The full-length α 1 subunit complementary DNA will also be useful as a dominant selectable marker for somatic cell genetic studies utilizing ouabain-sensitive cells.