Different behavior of l-afadin and Neurabin-II during the formation and destruction of cell-cell adherens junction

We have recently isolated two novel actin filament-binding proteins, l-afad in and neurabin-II and shown that they are localized at cell-cell adherens junction (AJ) in epithelial cells. We found here that l-afadin, neurabin-II , ZO-1, and E-cadherin showed similar and different behavior during the for mation and destruction of cell-cell AJ in MDCK cells. In MDCK cells, the ac cumulation of both l-afadin and E-cadherin, but not that of ZO-1, changed i n parallel depending on Rac small G protein activity. Dissociation of MDCK cells by culturing the cells at 2 mu M Ca2+ caused rapid endocytosis of E-c adherin, but not that of l-afadin or ZO-1, Addition of phorbol 12-myristate 13-acetate to these dissociated cells formed a tight junction-like structu re where ZO-1 and l-afadin, but not neurabin-II or E-cadherin, accumulated. We furthermore found that, in nonepithelial EL cells, which expressed E-ca dherin and attached to each other, l-afadin, neurabin-II, ZO-1 and E-cadher in were all localized at AJ. In cadherin-deficient L cells, l-afadin was ma inly localized at cell-cell contact sites, but ZO-1 was mainly localized at the tip area of cell processes. Neurabin-II did not accumulate at the plas ma membrane area. Neither l-afadin nor neurabin-II significantly interacted with alpha-, beta-catenin, E-cadherin, ZO-1 or occludin.