Amplification of DNA sequences coding for the Na,K-ATPase alpha-subunit in ouabain-resistant C+ cells.

J. R. Emanuel, S. Garetz, J. Schneider, J. F. Ash, E. J. Benz, R. Levenson

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We have studied the mechanism of cellular resistance to cardiac glycosides in C+ cells. C+ cells were resistant to ouabain and overproduced plasma membrane-bound Na,K-ATPase relative to parental HeLa cells. Overexpression of Na,K-ATPase in C+ cells correlated with increased ATPase mRNA levels and amplification (approximately 100 times) of the ATPase gene. Growth of C+ cells in ouabain-free medium resulted in a marked decline in ATPase mRNA and DNA levels. However, when cells were reexposed to ouabain, they proliferated and ATPase mRNA and DNA sequences were reamplified. Restriction analysis of C+ and other human DNA samples revealed the occurrence of rearrangements in the region of the Na,K-ATPase gene in C+ cells. Furthermore, C+ cells expressed an ATPase mRNA species not found in HeLa cells. These results suggest that amplification of the gene coding for Na,K-ATPase results in overproduction of Na,K-ATPase polypeptides. Amplification of the ATPase gene or the expression of new ATPase mRNA sequences or both may also be responsible for acquisition of the ouabain-resistant phenotype.

Original languageEnglish (US)
Pages (from-to)2476-2481
Number of pages6
JournalMolecular and Cellular Biology
Volume6
Issue number7
DOIs
StatePublished - Jan 1 1986

Fingerprint

Ouabain
Adenosine Triphosphatases
Messenger RNA
HeLa Cells
Cardiac Glycosides
Gene Amplification
DNA
sodium-translocating ATPase
Genes
Cell Membrane
Phenotype
Gene Expression
Peptides
Growth

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Amplification of DNA sequences coding for the Na,K-ATPase alpha-subunit in ouabain-resistant C+ cells. / Emanuel, J. R.; Garetz, S.; Schneider, J.; Ash, J. F.; Benz, E. J.; Levenson, R.

In: Molecular and Cellular Biology, Vol. 6, No. 7, 01.01.1986, p. 2476-2481.

Research output: Contribution to journalArticle

Emanuel, J. R. ; Garetz, S. ; Schneider, J. ; Ash, J. F. ; Benz, E. J. ; Levenson, R. / Amplification of DNA sequences coding for the Na,K-ATPase alpha-subunit in ouabain-resistant C+ cells. In: Molecular and Cellular Biology. 1986 ; Vol. 6, No. 7. pp. 2476-2481.
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