Role of actin in regulation of epithelial sodium channels by CFTR.

Academic Article

Abstract

  • Cystic fibrosis (CF) airway epithelia exhibit enhanced Na+ reabsorption in parallel with diminished Cl- secretion. We tested the hypothesis that actin plays a role in the regulation of a cloned epithelial Na+ channel (ENaC) by the cystic fibrosis transmembrane conductance regulator (CFTR). We found that immunopurified bovine tracheal CFTR coreconstituted into a planar lipid bilayer with alpha,beta,gamma-rat ENaC (rENaC) decreased single-channel open probability (Po) of rENaC in the presence of actin by over 60%, a significantly greater effect than was observed in the absence of actin (approximately 20%). In the presence of actin, protein kinase A plus ATP activated both CFTR and rENaC, but CFTR was activated in a sustained manner, whereas the activation of rENaC was transitory. ATP alone could also activate ENaC transiently in the presence ofactin but had no effect on CFTR. Stabilizing short actin filaments at a fixed length with gelsolin (at a ratio to actin of 2:1) produced a sustained activation of alpha,beta,gamma-rENaC in both the presence or absence of CFTR. Gelsolin alone (i.e., in the absence of actin) had no effect on the conductance or Po of either CFTR or rENaC. We have also found that short actin filaments produced their modulatory action on alpha-rENaC independent of the presence of the beta- or gamma-rENaC subunits. In contrast, CFTR did not affect any properties of the channel formed by alpha-rENaC alone, i.e., in the absence of beta- or gamma-rENaC. These results indicate that CFTR can directly downregulate single Na+ channel activity, which may account for the observed differences between Na+ transport in normal and CF-affected airway epithelia. Moreover, the presence of actin confers an enhanced modulatory ability of CFTR on Na+ channels.
  • Keywords

  • Actins, Adenosine Triphosphate, Animals, Cattle, Cyclic AMP-Dependent Protein Kinases, Cystic Fibrosis Transmembrane Conductance Regulator, Epithelium, Gelsolin, Isomerism, Lipid Bilayers, Rabbits, Rats, Respiratory System, Sodium Channels
  • Digital Object Identifier (doi)

    Author List

  • Ismailov II; Berdiev BK; Shlyonsky VG; Fuller CM; Prat AG; Jovov B; Cantiello HF; Ausiello DA; Benos DJ
  • Start Page

  • C1077
  • End Page

  • C1086
  • Volume

  • 272
  • Issue

  • 4 Pt 1