Expression of an ouabain-resistant Na,K-ATPase in CV-1 cells after transfection with a cDNA encoding the rat Na,K-ATPase alpha 1 subunit.

J. R. Emanuel, J. Schulz, X. M. Zhou, R. B. Kent, D. Housman, L. Cantley, Robert Levenson

Research output: Contribution to journalArticle

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Abstract

We have used a gene transfer system to investigate the relationship between expression of the rat Na,K-ATPase alpha 1 subunit gene and ouabain-resistant Na,K-ATPase activity. A cDNA clone encoding the entire rat Na,K-ATPase alpha 1 subunit was inserted into the expression vector pSV2neo. This construct (pSV2 alpha 1) conferred resistance to 100 microM ouabain to ouabain-sensitive CV-1 cells. Hybridization analysis of transfected clones revealed the presence of both rat-specific and endogenous Na,K-ATPase alpha 1 subunit DNA and mRNA sequences. A single form of highly ouabain-sensitive 86Rb+ uptake was detected in CV-1 cells, whereas two distinct classes of ouabain-inhibitable uptake were observed in transfectants. One class exhibited the high ouabain sensitivity of the endogenous monkey Na,K-ATPase, while the second class showed the reduced ouabain sensitivity characteristic of the rodent renal Na,K-ATPase. Examination of the ouabain-sensitive, sodium-dependent ATPase activity of the transfectants also revealed a low affinity component of Na,K-ATPase activity characteristic of the rodent kidney enzyme. These results suggest that expression of the rat alpha 1 subunit gene is directly responsible for ouabain-resistant Na,K-ATPase activity in transfected CV-1 cells.

Original languageEnglish (US)
Pages (from-to)7726-7733
Number of pages8
JournalThe Journal of biological chemistry
Volume263
Issue number16
StatePublished - Jan 1 1988

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Ouabain
Transfection
Adenosine Triphosphatases
Rats
Complementary DNA
Rodentia
Clone Cells
Genes
Gene transfer
Kidney
1,2-diamino-1,2-N,N'-carbonyl-1,2-dideoxyglucose hydrate
sodium-translocating ATPase
Haplorhini
Sodium
Messenger RNA
DNA
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Emanuel, J. R. ; Schulz, J. ; Zhou, X. M. ; Kent, R. B. ; Housman, D. ; Cantley, L. ; Levenson, Robert. / Expression of an ouabain-resistant Na,K-ATPase in CV-1 cells after transfection with a cDNA encoding the rat Na,K-ATPase alpha 1 subunit. In: The Journal of biological chemistry. 1988 ; Vol. 263, No. 16. pp. 7726-7733.
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Expression of an ouabain-resistant Na,K-ATPase in CV-1 cells after transfection with a cDNA encoding the rat Na,K-ATPase alpha 1 subunit. / Emanuel, J. R.; Schulz, J.; Zhou, X. M.; Kent, R. B.; Housman, D.; Cantley, L.; Levenson, Robert.

In: The Journal of biological chemistry, Vol. 263, No. 16, 01.01.1988, p. 7726-7733.

Research output: Contribution to journalArticle

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AU - Emanuel, J. R.

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