alpha-catenin-independent recruitment of ZO-1 to nectin-based cell-cell adhesion sites through afadin

ZO-1 is an actin filament (F-actin)-binding protein that localizes to tight junctions and connects claudin to the actin cytoskeleton in epithelial cel ls. In nonepithelial cells that have no tight junctions, ZO-1 localizes to adherens junctions (AJs) and may connect cadherin to the actin cytoskeleton indirectly through beta- and alpha -catenins as one of many F-actin-bindin g proteins. Nectin is an immunoglobulin-like adhesion molecule that localiz es to AJs and is associated with the actin cytoskeleton through afadin, an F-actin-binding protein. Ponsin is an afadin- and vinculin-binding protein that also localizes to AJs. The nectin-afadin complex has a potency to recr uit the E-cadherin-beta -catenin complex through alpha -catenin in a manner independent of ponsin. By the use of cadherin-deficient L cell lines stabl y expressing various components of the cadherin-catenin and nectin-afadin s ystems, and alpha -catenin- deficient F9 cell lines, we examined here wheth er nectin recruits ZO-1 to nectin-based cell-cell adhesion sites. Nectin sh owed a potency to recruit not only alpha -catenin but also ZO-1 to nectin-b ased cell-cell adhesion sites. This recruitment of ZO-1 was dependent on af adin but independent of alpha -catenin and ponsin. These results indicate t hat ZO-1 localizes to cadherin-based AJs through interactions not only with alpha -catenin but also with the nectin-afadin system.