Involvement of the Annexin II-S100A10 Complex in the Formation of E-cadherin-based Adherens Junctions in Madin-Darby Canine Kidney Cells

Abstract

E-cadherin and nectins are major cell-cell adhesion molecules at adherens junctions (AJs) in epithelial cells. When Madin-Darby canine kidney (MDCK) cells stably expressing nectin-1 (nectin-1-MDCK cells) are cultured at normal Ca(2+), E-cadherin and nectin-1 are concentrated at the cell-cell contact sites. When these cells are cultured at low Ca(2+), E-cadherin disappears from the cell-cell contact sites, but nectin-1 persists there. When these cells are re-cultured at normal Ca(2+), E-cadherin is recruited to the nectin-based cell-cell contact sites. We found here that this recruitment was dependent on protein synthesis, because a protein synthesis inhibitor, cycloheximide, prevented the accumulation of E-cadherin. When nectin-1-MDCK cells, precultured at low Ca(2+) in the presence of a proteasome inhibitor, ALLN (N-acetyl-Leu-Leu-norleucinal), were re-cultured at normal Ca(2+), E-cadherin was recruited to the nectin-based cell-cell contact sites but the level of E-cadherin was reduced. Similar results were obtained when wild-type MDCK cells were used instead of nectin-1-MDCK cells. These results suggest that degradation of one or more protein factors and de novo synthesis of the same or different protein factor(s) are needed for the formation of the E-cadherin-based AJs. We biochemically identified the annexin II-S100A10 complex as such a candidate. Depletion of plasma membrane cholesterol, which abolished the localization of the annexin II-S100A10 complex at the plasma membrane, inhibited the re-concentration of E-cadherin at the nectin-based cell-cell contact sites in the Ca(2+) switch experiment. Knockdown of annexin II by RNA interference also inhibited the re-concentration of E-cadherin. These results indicate that the annexin II-S100A10 complex is involved in the formation of the E-cadherin-based AJs in MDCK cells.

Regulation of epithelial cell polarity by PAR-3 depends on Girdin transcription and Girdin–Gαi3 signaling

“…It is possible that, by coordinating with the tiny amount of residual PAR-3 in PAR-3 kd cells, overexpressed Girdin can overcome the polarity defects caused by PAR-3 knockdown. Several early studies have demonstrated that increased synthesis of regulatory proteins is required for the establishment of cell-cell junctions (Griepp et al, 1983;Sang et al, 1980;Yamada et al, 2005). Our findings provide additional insight into this mechanism.…”

Tyrosine phosphorylation of annexin A2 regulates Rho-mediated actin rearrangement and cell adhesion

“…In epithelial cell layers, annexin A2 was shown to regulate cell spreading and wound closure in line with an increased expression of annexin A2 in migrating versus stationery epithelial cells (Babbin et al, 2007). Furthermore, in endothelial cells the reduction of annexin A2 plasma membrane levels by cholesterol depletion or RNAi inhibited the recruitment of E-cadherin and thereby cell contact formation, which itself negatively regulates migration (Yamada et al, 2005). It will be interesting to learn whether the effect of annexin A2 on cell contact formation is also regulated by its tyrosine phosphorylation, which has been established here as a molecular switch transforming cellular actin from a stationary to a more migratory phenotype.…”

Anchoring Junctions As Drug Targets: Role in Contraceptive Development

“…Nectins also regulate cytoskeletal dynamics via F-actin binding proteins that interact directly with nectin [e.g., afadin, ␣-catenin, ␣-actinin, vinculin, annexin II and IQGAP Miyoshi and Takai, 2007)] (Fig. 1), and establishment of nectin-based adherens junctions required annexin II and IQGAP in MDCK cells (Katata et al, 2003;Yamada et al, 2005aYamada et al, , 2006a. Furthermore, nectin-afadin can also recruit tight junction proteins such as claudin, occludin, JAM-A, and ZO-1 to the junctional complex (Yokoyama et al, 2001;Fukuhara et al, 2002;Takai and Nakanishi, 2003;Sato et al, 2006), suggesting the existence of cross talk between different junction types.…”