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Regulation of Na/K-ATPase β1-subunit gene expression by ouabain and other hypertrophic stimuli in neonatal rat cardiac myocytes

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Abstract

Partial inhibition of Na/K-ATPase by ouabain causes hypertrophic growth and regulates several early and late response genes, including that of Na/K-ATPase α3 subunit, in cultured neonatal rat cardiac myocytes. The aim of this work was to determine whether ouabain and other hypertrophic stimuli affect Na/K-ATPase β1 subunit gene expression. When myocytes were exposed to non-toxic concentrations of ouabain, ouabain increased β1 subunit mRNA in a dose- and time-dependent manner. Like the α3 gene, β1 mRNA was also regulated by several other well-known hypertrophic stimuli including phenylephrine, a phorbol ester, endothelin-1, and insulin-like growth factor, suggesting involvement of growth signals in regulation of β1 expression. Ouabain failed to increase β1 subunit mRNA in the presence of actinomycin D. Using a luciferase reporter gene that is directed by the 5′-flanking region of the β1 subunit gene, transient transfection assay showed that ouabain augmented the expression of luciferase. These data support the proposition that ouabain regulates the β1 subunit through a transcriptional mechanism. The effect of ouabain on β1 subunit induction, like that on α3 repression, was dependent on extracellular Ca2+ and on calmodulin. Inhibitions of PKC, Ras, and MEK, however, had different quantitive effects on ouabain-induced regulations of β1 and α3 subunits. The findings show that partial inhibition of Na/K-ATPase activates multiple signaling pathways that regulate growth-related genes, including those of two subunit isoforms of Na/K-ATPase, in a gene-specific manner.

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Authors and Affiliations

  1. Department of Pharmacology, Medical College of Ohio, Toleda, OH, 43614, USA

    Peter Kometiani, Jiang Tian & Ziad Nabih

  2. Department of Biochemistry, Health Science Center, State University of New York, Brooklyn, NY, 11203, USA

    Gregory Gick

  3. Department of Pharmacology, Medical College of Ohio, Toleda, OH, 43614, USA

    Zijian Xie

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  1. Peter Kometiani
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  2. Jiang Tian
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  4. Gregory Gick
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  5. Zijian Xie
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Kometiani, P., Tian, J., Nabih, Z. et al. Regulation of Na/K-ATPase β1-subunit gene expression by ouabain and other hypertrophic stimuli in neonatal rat cardiac myocytes. Mol Cell Biochem 215, 65–72 (2000). https://doi.org/10.1023/A:1026503319387

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