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Papers In Press, published online ahead of print August 31, 2004
Department of Medicine, Physiology and Pediatrics, University of Minnesota, Minneapolis, MN 55455
Corresponding Author: ingba001{at}umn.edu
We previously reported that thyroid hormone, 3,3’,5-triiodo-L-thyronine (T3), increased Na,K-ATPase activity of adult rat alveolar epithelial cells in a transcription-independent manner via increased cell surface expression of the 1 and 1 subunits of Na,K-ATPase. Now we sought to identify signaling molecules necessary for T3 stimulation of Na,K-ATPase activity in alveolar epithelial cells. While protein kinase A inhibitor H-8 and protein kinase C inhibitor bisindolymaleimide did not block the T3-induced increase in Na,K-ATPase activity, two inhibitors of PI3K, wortmannin and Ly294002, and two Src kinase inhibitors, PP1 and PP2, blocked the T3-induced Na,K-ATPase activity. T3 stimulated the activity of PI3K as measured by PI(3)P. T3 also stimulated the serine-473 phosphorylation of the PI3K downstream molecule PKB/Akt in a dose-dependent manner. Transient expression of a constitutively active mutant of the PI3-kinase catalytic subunit p110 augmented Na,K-ATPase activity and increased the amount of cell surface Na,K-ATPase 1 subunit protein. T3 also stimulated Src family kinase activity. T ransient expression of a constitutively active Src kinase increased Na,K-ATPase activity, PI3K activity and phosphorylation of PKB/Akt at serine-473. PP1 or PP2 blocked T3-stimulated PKB/Akt phosphorylation at serine-473 and PI3K activity that activated by an active mutant of Src, however, wortmannin did not inhibit the T3-stimulated Src kinase activity. Although PP1 and wortmannin abolished the increase in Na,K-ATPase activity induced by the active mutant of Src, PP1 did not inhibit the active mutant of PI3K-upregulated Na,K-ATPase activity. In summary, T3 stimulates the PI3K/PKB pathway via the Src family of tyrosine kinases and activation of both the Src family kinases and PI3-kinase is required for the T3-induced stimulation of Na,K-ATPase activity and its cell surface expression in adult rat alveolar epithelial cells.
J. Biol. Chem, 10.1074/jbc.M405497200
Submitted on May 17, 2004
Revised on August 31, 2004
Accepted on August 31, 2004
3,3',5-triiodo-L-thyronine (T3) upregulation of Na,K-ATPase activity and cell surface expression in alveolar epithelial cells is Src kinase and PI3K dependent
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