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

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

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    35 Citations (Scopus)

    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 beta- and gamma- 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 (alpha beta(S631A)gamma(T599A)) 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 (alpha beta(S631A)gamma(T599A)-ENaC. Phosphorylation by CK2 is essential for ENaC activation, and to a lesser degree, it also controls membrane expression of alpha beta gamma-ENaC. Channels lacking the Nedd4-2 binding motif in beta-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 beta-ENaC is targeted by this essential, conserved pleiotropic kinase that directs its constitutive activity toward many cellular protein complexes.

    Original languageEnglish
    Pages (from-to)13225-13232
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume283
    Issue number19
    DOIs
    Publication statusPublished - 9 May 2008

    Keywords

    • TRANSMEMBRANE CONDUCTANCE REGULATOR
    • CELL-SURFACE EXPRESSION
    • SODIUM-CHANNEL
    • OPEN PROBABILITY
    • DOWN-REGULATION
    • ALPHA-SUBUNIT
    • BETA-SUBUNIT
    • PHOSPHORYLATION
    • ENAC
    • SERUM

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