Regulation of the epithelial Na+ channel by the protein kinase CK2

Tanja Bachhuber, Joana Almaça, Fadi Aldehni, Anil Mehta, Margarida D. Amaral, Rainer Schreiber, Karl Kunzelmann

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33 Scopus citations

Abstract

CK2 is a ubiquitous, pleiotropic, and constitutively active Ser/Thr protein kinase that controls protein expression, cell signaling, and ion channel activity. Phosphorylation sites for CK2 are located in the C terminus of both β- and γ-subunits of the epithelial Na+ channel (ENaC). We examined the role of CK2 on the regulation of both endogenous ENaC in native murine epithelia and in Xenopus oocytes expressing rENaC. In Ussing chamber experiments with mouse airways, colon, and cultured M1-collecting duct cells, amiloride-sensitive Na+ transport was inhibited dose-dependently by the selective CK2 inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB). In oocytes, ENaC currents were also inhibited by TBB and by the structurally unrelated inhibitors heparin and poly(E:Y). Expression of a trimeric channel lacking both CK2 sites (αβS631AγT599A) produced a largely attenuated amiloride-sensitive whole cell conductance and rendered the mutant channel insensitive to CK2. In Xenopus oocytes, CK2 was translocated to the cell membrane upon expression of wt-ENaC but not of αβ S631AγT599A-ENaC. Phosphorylation by CK2 is essential for ENaC activation, and to a lesser degree, it also controls membrane expression of αβγ-ENaC. Channels lacking the Nedd4-2 binding motif in β-ENaC (R561X, Y618A) no longer required the CK2 site for channel activity and siRNA-knockdown of Nedd4-2 eliminated the effects of TBB. This implies a role for CK2 in inhibiting the Nedd4-2 pathway. We propose that the C terminus of β-ENaC is targeted by this essential, conserved pleiotropic kinase that directs its constitutive activity toward many cellular protein complexes.

Original languageEnglish (US)
Pages (from-to)13225-13232
Number of pages8
JournalJournal of Biological Chemistry
Volume283
Issue number19
DOIs
StatePublished - May 9 2008
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Bachhuber, T., Almaça, J., Aldehni, F., Mehta, A., Amaral, M. D., Schreiber, R., & Kunzelmann, K. (2008). Regulation of the epithelial Na+ channel by the protein kinase CK2. Journal of Biological Chemistry, 283(19), 13225-13232. https://doi.org/10.1074/jbc.M704532200