Characterization of the association of connexins and ZO-1 in the lens

ZO-1 (Zona Occludens protein 1) has previously been shown to bind Cx43alpha1. This interaction involves the most C-terminal residues of Cx43alpha1 and the second PDZ-domain of ZO-1. The biological significance of this interaction is not well understood. The similarity of the C-terminal residues of the lens connexins Cx46alpha3 and Cx50alpha8 to Cx43alpha1 prompted us to examine if ZO-1 is expressed in the lens, and if ZO-1 interacts with lens connexins. A high level of ZO-1 expression was detected in the mouse lens. Lens connexins were shown to co-immunoprecipitate with ZO-1, and the interaction was found to involve similar domains as those previously demonstrated for the Cx43alpha1/ZO-1 interaction (Nielsen et al. manuscript in preparation). Futhermore, transient expression of Cx46alpha3 and Cx50alpha8 in cell culture showed colocalization of gap junction plaques with ZO-1, further suggesting that lens connexins interact with ZO-1. Sequence comparison suggests that a large number of connexins of the alpha subclass may interact with ZO-1. Using the lens as a system to study connexin/ZO-1 interactions may further our understanding of their biological significance in the lens, as well as in other organs.