Abstract
We investigated the possible functional‐ and physical protein‐interactions between two airway Cl^−^ channels, SLC26A9 and CFTR. Bronchial CFBE41o‐ cell lines expressing CFTR~WT~ or CFTR~ΔF508~ were transduced with SLC26A9. Immunoblots identified a migrating band corresponding to SLC26A9 present in whole‐cell lysates as on apical membrane of cells grown on polarized filters. CFTR levels were increased by the presence of SLC26A9 in both CFTR~WT~ and CFTR~ΔF508~ cell lines. In CFBE41o‐ cells and CFBE41o‐/CFTR~WT~ cells transduced with SLC26A9, currents associated to the protein expression were not detected. However, the forskolin (FK)‐stimulated currents were enhanced in SLC26A9‐transduced cells compared to control cells. Therefore, the presence of SLC26A9 resulted in an increase in CFTR activity (same % of CFTR~(inh)‐172~ or GlyH‐101 inhibition in both groups). In CFBE41o‐/CFTR~ΔF508~ cells transduced with SLC26A9 (at 27°C), a current associated to the protein expression was also lacking. FK‐stimulated currents and level of CFTR~(inh)‐172~ inhibition were not different in both groups. The presence of SLC26A9 in Xenopus oocytes expressing CFTR also enhanced the FK‐stimulated currents as compared to oocytes expressing CFTR alone. This stimulation was mostly linked to CFTR. An enhancement of FK‐stimulated currents was not found in oocytes co‐expressing SLC26A9 and CFTR~ΔF508~. In conclusion, in both protein expression systems used, SLC26A9 stimulates CFTR activity but not that of CFTR~ΔF508~. Our co‐immunoprecipitation studies demonstrate a physical interaction between both anion channels. We propose as an alternative hypothesis (not exclusive) to the known SLC26A9‐STAS domain/CFTR interaction, that SLC26A9 favors the biogenesis and/or stabilization of CFTR, leading to stimulated currents. J. Cell. Physiol. 226: 212–223, 2010. © 2010 Wiley‐Liss, Inc.