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Cadherins, Ca 2+ -dependent adhesion molecules, are crucial for cell-cell junctions and remodeling. Cadherins form inter-junctional lattices by the formation of both  cis  and  trans  dimers. Here, we directly visualize and quantify the spatiotemporal dynamics of wild-type and dimer mutant N-cadherin interactions using time-lapse imaging of junction assembly, disassembly and a FRET reporter to assess Ca 2+ -dependent interactions. A  trans  dimer mutant (W2A) and a  cis  mutant (V81D/V174D) exhibited an increased Ca 2+ -sensitivity for the disassembly of  trans  dimers compared to the WT, while another mutant (R14E) was insensitive to Ca 2+ -chelation. Time-lapse imaging of junction assembly and disassembly, monitored in 2D and 3D (using cellular spheroids), revealed kinetic differences in the different mutants as well as different behaviors in the 2D and 3D environment. Taken together, these data provide new insights into the role that the  cis  and  trans  dimers play in the dynamic interactions of cadherins.

Role of N-Cadherin cis and trans Interfaces in the Dynamics of Adherens Junctions in Living Cells