CFTR Controls the Activity of NF-kappa B by Enhancing the Degradation of TRADD

Background/Aims: Chronic lung infection in cystic fibrosis leads to an inflammatory response that persists because of the chronic presence of bacteria and ultimately leads to a catastrophic failure of lung function. Methods: We use a combination of biochemistry, cell and molecular biology to study the interaction of TRADD, a key adaptor molecule in TNF alpha signaling, with CFTR in the regulation of NF kappa B. Results: We show that Wt CFTR binds to and colocalizes with TRADD. TRADD is a key signaling intermediate connecting TNFa with activation of NF kappa B. By contrast,Delta F508 CFTR does not bind to TRADD. NF-kappa B activation is higher in CFBE expressing Delta F508 CFTR than in cells expressing Wt CFTR. However, this differential effect is abolished when TRADD levels are knocked down. Transfecting Wt CFTR into CFBE cells reduces NF kappa B activity. However the reduction is abolished by the CFTR chloride transport inhibitor-172. Consistently, transfecting in the correctly trafficked CFTR conduction mutants G551D or S341A also fail to reduce NF kappa B activity. Thus CFTR must be functional if it is to regulate NF-kappa B activity. We also found that TNFa produced a greater increase in NF-kappa B activity in CFBE cells than in the same cell when Wt CFTR-corrected. Consistently, the effect is also abolished when TRADD is knocked down by shRNA. Thus, Wt CFTR control of TRADD modulates the physiological activation of NF-kappa B by TNFa. Based on studies with proteosomal and lysosomal inhibitors, the mechanism by which Wt CFTR, but not Delta F508 CFTR, suppresses TRADD is by lysosomal degradation. Conclusion: We have uncovered a novel mechanism whereby Wt CFTR regulates TNFa signaling by enhancing TRADD degradation. Thus by reducing the levels of TRADD, Wt CFTR suppresses downstream proinflammatory NF kappa B signaling. By contrast, suppression of NF-kappa B activation fails in CF cells expressing Delta F508 CFTR. (C) 2016 The Author(s) Published by S. Karger AG, Basel