XB-ART-40027FASEB J. November 1, 2009; 23 (11): 3829-42.
Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species.
The mechanisms by which replicating influenza viruses decrease the expression and function of amiloride-sensitive epithelial sodium channels (ENaCs) have not been elucidated. We show that expression of M2, a transmembrane influenza protein, decreases ENaC membrane levels and amiloride-sensitive currents in both Xenopus oocytes, injected with human alpha-, beta-, and gamma-ENaCs, and human airway cells (H441 and A549), which express native ENaCs. Deletion of a 10-aa region within the M2 C terminus prevented 70% of this effect. The M2 ENaC down-regulation occurred at normal pH and was prevented by MG-132, a proteasome and lysosome inhibitor. M2 had no effect on Liddle ENaCs, which have decreased affinity for Nedd4-2. H441 and A549 cells transfected with M2 showed higher levels of reactive oxygen species, as shown by the activation of redox-sensitive dyes. Pretreatment with glutathione ester, which increases intracellular reduced thiol concentrations, or protein kinase C (PKC) inhibitors prevented the deleterious effects of M2 on ENaCs. The data suggest that M2 protein increases steady-state concentrations of reactive oxygen intermediates that simulate PKC and decrease ENaCs by enhancing endocytosis and its subsequent destruction by the proteasome. These novel findings suggest a mechanism for the influenza-induced rhinorrhea and life-threatening alveolar edema in humans.
PubMed ID: 19596899
PMC ID: PMC2775009
Article link: FASEB J.
Grant support: 1U54ES017218 NIEHS NIH HHS , 5P30DK072482 NIDDK NIH HHS , 5U01ES015676 NIEHS NIH HHS , AI071393 NIAID NIH HHS , HL031197 NHLBI NIH HHS , R01 AI071393 NIAID NIH HHS , P30 DK072482 NIDDK NIH HHS , R37 HL031197 NHLBI NIH HHS , U54 ES017218 NIEHS NIH HHS , R01 HL031197 NHLBI NIH HHS , U01 ES015676 NIEHS NIH HHS
Genes referenced: nedd4 nedd4l