An optochemical tool for light-induced dissociation of adherens junctions to control mechanical coupling between cells

Dirk Ollech,Elisabetta Ada Cavalcanti-Adam,Tim Pflästerer,Roberto Mayor,Joachim Pius Spatz,Chiara Zambarda,Philippe Marcq,Adam Shellard,Richard Wombacher

doi:10.1038/s41467-020-14390-1

Abstract

The cadherin-catenin complex at adherens junctions (AJs) is essential for the formation of cell-cell adhesion and epithelium integrity; however, studying the dynamic regulation of AJs at high spatio-temporal resolution remains challenging. Here we present an optochemical tool which allows reconstitution of AJs by chemical dimerization of the force bearing structures and their precise light-induced dissociation. For the dimerization, we reconstitute acto-myosin connection of a tailless E-cadherin by two ways: direct recruitment of α-catenin, and linking its cytosolic tail to the transmembrane domain. Our approach enables a specific ON-OFF switch for mechanical coupling between cells that can be controlled spatially on subcellular or tissue scale via photocleavage. The combination with cell migration analysis and traction force microscopy shows a wide-range of applicability and confirms the mechanical contribution of the reconstituted AJs. Remarkably, in vivo our tool is able to control structural and functional integrity of the epidermal layer in developing Xenopus embryos.

Highlights

The cadherin-catenin complex at adherens junctions (AJs) is essential for the formation of cell-cell adhesion and epithelium integrity; studying the dynamic regulation of AJs at high spatio-temporal resolution remains challenging
We present LInDA (Light-Induced Dissociation of Adherens Junctions), an optochemical tool based on photocleavable dimerizers[13,14,15,16] for the precise control of E-cadherin-mediated AJs dynamics (Fig. 1a)
We developed two setups to control with LInDA the assembly and disassembly of AJs in epithelial cells which are lacking these structures. (i) To establish the E-cadherin/α-catenin linkage we replaced the cytosolic tail domain of E-cadherin with Halo tag[17] (E-cadherin-Δcyto-Halo) and the N-terminal domain of α-catenin with SNAP tag[18] (SNAP-ΔN-α-catenin) (Fig. 1d and Supplementary Fig. 1a–d). (ii) To reconstitute the E-cadherin protein, we combined the expression of E-cadherin-Δcyto-Halo with the expression of the cytosolic tail of E-cadherin presenting a DHFR tag[19] (DHFR-cyto) (Fig. 1e and Supplementary Fig. b, e–g)

Summary

Introduction

The cadherin-catenin complex at adherens junctions (AJs) is essential for the formation of cell-cell adhesion and epithelium integrity; studying the dynamic regulation of AJs at high spatio-temporal resolution remains challenging. Dynamic regulation of AJ formation and dissociation is essential for homeostasis and functional integrity of epithelial tissues These mechanisms have been studied using constitutive or inducible knockout strategies[9], or by preventing E-cadherin interaction either using antibodies[10,11] or Ca2+ depletion[11,12]. These tools gave remarkable insights into AJ formation dynamics, they are limited in temporal resolution and lack spatial control. We present LInDA (Light-Induced Dissociation of Adherens Junctions), an optochemical tool based on photocleavable dimerizers[13,14,15,16] for the precise control of E-cadherin-mediated AJs dynamics (Fig. 1a). We developed two setups to control with LInDA the assembly and disassembly of AJs in epithelial cells which are lacking these structures (either from knockout or due to transformation-associated loss in protein expression). (i) To establish the E-cadherin/α-catenin linkage we replaced the cytosolic tail domain of E-cadherin with Halo tag[17] (E-cadherin-Δcyto-Halo) and the N-terminal domain of α-catenin with SNAP tag[18] (SNAP-ΔN-α-catenin) (Fig. 1d and Supplementary Fig. 1a–d). (ii) To reconstitute the E-cadherin protein, we combined the expression of E-cadherin-Δcyto-Halo with the expression of the cytosolic tail of E-cadherin presenting a DHFR tag[19] (DHFR-cyto) (Fig. 1e and Supplementary Fig. b, e–g)

Methods

Results

Conclusion