scholarly journals The formation of ordered nanoclusters controls cadherin anchoring to actin and cell–cell contact fluidity

2015 ◽  
Vol 210 (2) ◽  
pp. 333-346 ◽  
Author(s):  
Pierre-Olivier Strale ◽  
Laurence Duchesne ◽  
Grégoire Peyret ◽  
Lorraine Montel ◽  
Thao Nguyen ◽  
...  
Keyword(s):  
Adherens Junction ◽  
Intercellular Junctions ◽  
Cell Contact ◽  
Collective Cell Migration ◽  
Contact Stability ◽  
Junction Formation ◽  
Mediate Cell ◽  
The Stability ◽  
E Cadherin ◽  
Cell Cell

Oligomerization of cadherins could provide the stability to ensure tissue cohesion. Cadherins mediate cell–cell adhesion by forming trans-interactions. They form cis-interactions whose role could be essential to stabilize intercellular junctions by shifting cadherin clusters from a fluid to an ordered phase. However, no evidence has been provided so far for cadherin oligomerization in cellulo and for its impact on cell–cell contact stability. Visualizing single cadherins within cell membrane at a nanometric resolution, we show that E-cadherins arrange in ordered clusters, providing the first demonstration of the existence of oligomeric cadherins at cell–cell contacts. Studying the consequences of the disruption of the cis-interface, we show that it is not essential for adherens junction formation. Its disruption, however, increased the mobility of junctional E-cadherin. This destabilization strongly affected E-cadherin anchoring to actin and cell–cell rearrangement during collective cell migration, indicating that the formation of oligomeric clusters controls the anchoring of cadherin to actin and cell–cell contact fluidity.

scholarly journals Force-dependent binding of vinculin to α-catenin regulates cell–cell contact stability and collective cell behavior

2018 ◽  
Vol 29 (4) ◽  
pp. 380-388 ◽  
Author(s):  
Rima Seddiki ◽  
Gautham Hari Narayana Sankara Narayana ◽  
Pierre-Olivier Strale ◽  
Hayri Emrah Balcioglu ◽  
Grégoire Peyret ◽  
...  
Keyword(s):  
Mechanical Load ◽  
Intercellular Junctions ◽  
Magnetic Tweezers ◽  
Contact Dynamics ◽  
Fine Tuning ◽  
Cell Contact ◽  
Collective Cell Migration ◽  
Mechanical Coupling ◽  
Contact Stability ◽  
Cell Cell

The shaping of a multicellular body and repair of adult tissues require fine-­tuning of cell adhesion, cell mechanics, and intercellular transmission of mechanical load. Adherens junctions (AJs) are the major intercellular junctions by which cells sense and exert mechanical force on each other. However, how AJs adapt to mechanical stress and how this adaptation contributes to cell–cell cohesion and eventually to tissue-scale dynamics and mechanics remains largely unknown. Here, by analyzing the tension-dependent recruitment of vinculin, α-catenin, and F-actin as a function of stiffness, as well as the dynamics of GFP-tagged wild-type and mutated α-catenins, altered for their binding capability to vinculin, we demonstrate that the force-dependent binding of vinculin stabilizes α-catenin and is responsible for AJ adaptation to force. Challenging cadherin complexes mechanical coupling with magnetic tweezers, and cell–cell cohesion during collective cell movements, further highlight that tension-dependent adaptation of AJs regulates cell–cell contact dynamics and coordinated collective cell migration. Altogether, these data demonstrate that the force-dependent α-catenin/vinculin interaction, manipulated here by mutagenesis and mechanical control, is a core regulator of AJ mechanics and long-range cell–cell interactions.

scholarly journals Biochemical and structural analysis of α-catenin in cell–cell contacts

2008 ◽  
Vol 36 (2) ◽  
pp. 141-147 ◽  
Author(s):  
Sabine Pokutta ◽  
Frauke Drees ◽  
Soichiro Yamada ◽  
W. James Nelson ◽  
William I. Weis
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Adherens Junction ◽  
Structural Data ◽  
Cell Contact ◽  
Related Protein ◽  
Imaging Studies ◽  
Actin Networks ◽  
Junction Formation ◽  
Cell Cell

Cadherins are transmembrane adhesion molecules that mediate homotypic cell–cell contact. In adherens junctions, the cytoplasmic domain of cadherins is functionally linked to the actin cytoskeleton through a series of proteins known as catenins. E-cadherin binds to β-catenin, which in turn binds to α-catenin to form a ternary complex. α-Catenin also binds to actin, and it was assumed previously that α-catenin links the cadherin–catenin complex to actin. However, biochemical, structural and live-cell imaging studies of the cadherin–catenin complex and its interaction with actin show that binding of β-catenin to α-catenin prevents the latter from binding to actin. Biochemical and structural data indicate that α-catenin acts as an allosteric protein whose conformation and activity changes depending on whether or not it is bound to β-catenin. Initial contacts between cells occur on dynamic lamellipodia formed by polymerization of branched actin networks, a process controlled by the Arp2/3 (actin-related protein 2/3) complex. α-Catenin can suppress the activity of Arp2/3 by competing for actin filaments. These findings lead to a model for adherens junction formation in which clustering of the cadherin–β-catenin complex recruits high levels of α-catenin that can suppress the Arp2/3 complex, leading to cessation of lamellipodial movement and formation of a stable contact. Thus α-catenin appears to play a central role in cell–cell contact formation.

scholarly journals EphA2 Engages Git1 to Suppress Arf6 Activity Modulating Epithelial Cell–Cell Contacts

2009 ◽  
Vol 20 (7) ◽  
pp. 1949-1959 ◽  
Author(s):  
Koichi Miura ◽  
Jin-Min Nam ◽  
Chie Kojima ◽  
Naoki Mochizuki ◽  
Hisataka Sabe
Keyword(s):  
Ion Concentration ◽  
Cell Contact ◽  
Interacting Protein ◽  
Cell Contacts ◽  
Extracellular Signal ◽  
Src Homology ◽  
Src Homology 2 Domain ◽  
Mediate Cell ◽  
E Cadherin ◽  
Cell Cell

ADP-ribosylation factor (Arf) 6 activity is crucially involved in the regulation of E-cadherin–based cell–cell adhesions. Erythropoietin-producing hepatocellular carcinoma (Eph)-family receptors recognize ligands, namely, ephrins, anchored to the membrane of apposing cells, and they mediate cell–cell contact-dependent events. Here, we found that Arf6 activity is down-regulated in Madin-Darby canine kidney cells, which is dependent on cell density and calcium ion concentration, and we provide evidence of a novel signaling pathway by which ligand-activated EphA2 suppresses Arf6 activity. This EphA2-mediated suppression of Arf6 activity was linked to the induction of cell compaction and polarization, but it was independent of the down-regulation of extracellular signal-regulated kinase 1/2 kinase activity. We show that G protein-coupled receptor kinase-interacting protein (Git) 1 and noncatalytic region of tyrosine kinase (Nck) 1 are involved in this pathway, in which ligand-activated EphA2, via its phosphorylated Tyr594, binds to the Src homology 2 domain of Nck1, and then via its Src homology 3 domain binds to the synaptic localizing domain of Git1 to suppress Arf6 activity. We propose a positive feedback loop in which E-cadherin–based cell–cell contacts enhance EphA-ephrinA signaling, which in turn down-regulates Arf6 activity to enhance E-cadherin–based cell–cell contacts as well as the apical-basal polarization of epithelial cells.

scholarly journals Plakophilin 3 mediates Rap1-dependent desmosome assembly and adherens junction maturation

2014 ◽  
Vol 25 (23) ◽  
pp. 3749-3764 ◽  
Author(s):  
Viktor Todorovic´ ◽  
Jennifer L. Koetsier ◽  
Lisa M. Godsel ◽  
Kathleen J. Green
Keyword(s):  
Initial Phase ◽  
Adherens Junction ◽  
Cell Contact ◽  
Functional Association ◽  
Plakophilin 2 ◽  
Plakophilin 3 ◽  
E Cadherin ◽  
Cell Cell

The pathways driving desmosome and adherens junction assembly are temporally and spatially coordinated, but how they are functionally coupled is poorly understood. Here we show that the Armadillo protein plakophilin 3 (Pkp3) mediates both desmosome assembly and E-cadherin maturation through Rap1 GTPase, thus functioning in a manner distinct from the closely related plakophilin 2 (Pkp2). Whereas Pkp2 and Pkp3 share the ability to mediate the initial phase of desmoplakin (DP) accumulation at sites of cell–cell contact, they play distinct roles in later steps: Pkp3 is required for assembly of a cytoplasmic population of DP-enriched junction precursors, whereas Pkp2 is required for transfer of the precursors to the membrane. Moreover, Pkp3 forms a complex with Rap1 GTPase, promoting its activation and facilitating desmosome assembly. We show further that Pkp3 deficiency causes disruption of an E-cadherin/Rap1 complex required for adherens junction sealing. These findings reveal Pkp3 as a coordinator of desmosome and adherens junction assembly and maturation through its functional association with Rap1.

scholarly journals Non-junctional Cadherin3 regulates cell migration and contact inhibition of locomotion via domain-dependent opposing regulations of Rac1

2019 ◽  
Author(s):  
Takehiko Ichikawa ◽  
Carsten Stuckenholz ◽  
Lance A. Davidson
Keyword(s):  
Cell Migration ◽  
Contact Inhibition ◽  
Cell Contact ◽  
Collective Cell Migration ◽  
Cell Contacts ◽  
Rac1 Activity ◽  
Classical Cadherins ◽  
The Stability ◽  
Cell Cell

AbstractClassical cadherins are well-known primary adhesion molecules responsible for physically connecting neighboring cells and signaling the cell-cell contact. Recent studies have suggested novel signaling roles for “non-junctional” cadherins (Niessen and Gottardi, 2008; Padmanabhan et al., 2017); however, the function of cadherin signaling independent of cell-cell contacts remains unknown. In this study, we used mesendoderm cells and tissues from gastrula stage Xenopus laevis embryos to demonstrate that extracellular and cytoplasmic cadherin domains regulate Rac1 in opposing directions in the absence of cell-cell contacts. Furthermore, we found that non-junctional cadherins regulate contact inhibition of locomotion (CIL) during gastrulation through alterations in the stability of the cytoskeleton. Live FRET imaging of Rac1 activity illustrated how non-junction cadherin3 (formerly C-cadherin) spatio-temporally regulates CIL. Our study provides novel insights into adhesion-independent signaling by cadherin3 and its role in regulating single and collective cell migration in vivo.

scholarly journals Cortical actin flow activates an alpha-catenin clutch to assemble adherens junctions

2021 ◽  
Author(s):  
Ivar Noordstra ◽  
Mario Diez Hermoso ◽  
Lilian Schimmel ◽  
Alexis Bonfim-Melo ◽  
Joseph Mathew Kalappurakkal ◽  
...  
Keyword(s):  
Adherens Junctions ◽  
Actin Binding ◽  
Cell Contact ◽  
Cortical Actin ◽  
Bond Behaviour ◽  
Catch Bond ◽  
Actin Cortex ◽  
Mediate Cell ◽  
E Cadherin ◽  
Cell Cell

Adherens junctions (AJs) fundamentally mediate cell-cell adhesion, yet the mechanisms that determine where or when AJs assemble are not understood. Here we reveal a mechanosensitive clutch that initiates AJ assembly. Before cell-cell contact, alpha-catenin couples surface E-cadherin complexes to retrograde flow of the actin cortex. Cortical flows with opposed orientations persist after contact, applying tension to alpha-catenin within trans-ligated cadherin complexes. Tension unfolds the alpha-catenin actin-binding domain (ABD), which is expected to mediate a catch bond with F-actin. However, catch bond behaviour is not sufficient for AJ assembly in a molecular clutch model. Instead, it is also necessary for the activated ABD to promote cis-clustering of E-cadherin molecules by bundling F-actin. Thus, this alpha-catenin clutch transduces the mechanical signal of cortical flow to assemble AJs.

scholarly journals Remodeling the cell surface distribution of membrane proteins during the development of epithelial cell polarity.

1992 ◽  
Vol 116 (4) ◽  
pp. 889-899 ◽  
Author(s):  
D A Wollner ◽  
K A Krzeminski ◽  
W J Nelson
Keyword(s):  
Epithelial Cell ◽  
Membrane Proteins ◽  
Cell Surface ◽  
Mdck Cells ◽  
Apical Cell ◽  
Cell Contact ◽  
Surface Polarity ◽  
Lateral Membrane ◽  
E Cadherin ◽  
Cell Cell

The development of polarized epithelial cells from unpolarized precursor cells follows induction of cell-cell contacts and requires resorting of proteins into different membrane domains. We show that in MDCK cells the distributions of two membrane proteins, Dg-1 and E-cadherin, become restricted to the basal-lateral membrane domain within 8 h of cell-cell contact. During this time, however, 60-80% of newly synthesized Dg-1 and E-cadherin is delivered directly to the forming apical membrane and then rapidly removed, while the remainder is delivered to the basal-lateral membrane and has a longer residence time. Direct delivery of greater than 95% of these proteins from the Golgi complex to the basal-lateral membrane occurs greater than 48 h later. In contrast, we show that two apical proteins are efficiently delivered and restricted to the apical cell surface within 2 h after cell-cell contact. These results provide insight into mechanisms involved in the development of epithelial cell surface polarity, and the establishment of protein sorting pathways in polarized cells.

scholarly journals Changes in E-cadherin rigidity sensing regulate cell adhesion

2017 ◽  
Vol 114 (29) ◽  
pp. E5835-E5844 ◽  
Author(s):  
Caitlin Collins ◽  
Aleksandra K. Denisin ◽  
Beth L. Pruitt ◽  
W. James Nelson
Keyword(s):  
Cell Adhesion ◽  
Signaling Molecules ◽  
Membrane Dynamics ◽  
Cell Contact ◽  
Adhesion Assay ◽  
Cell Periphery ◽  
Actin Organization ◽  
Rigidity Sensing ◽  
E Cadherin ◽  
Cell Cell

Mechanical cues are sensed and transduced by cell adhesion complexes to regulate diverse cell behaviors. Extracellular matrix (ECM) rigidity sensing by integrin adhesions has been well studied, but rigidity sensing by cadherins during cell adhesion is largely unexplored. Using mechanically tunable polyacrylamide (PA) gels functionalized with the extracellular domain of E-cadherin (Ecad-Fc), we showed that E-cadherin–dependent epithelial cell adhesion was sensitive to changes in PA gel elastic modulus that produced striking differences in cell morphology, actin organization, and membrane dynamics. Traction force microscopy (TFM) revealed that cells produced the greatest tractions at the cell periphery, where distinct types of actin-based membrane protrusions formed. Cells responded to substrate rigidity by reorganizing the distribution and size of high-traction-stress regions at the cell periphery. Differences in adhesion and protrusion dynamics were mediated by balancing the activities of specific signaling molecules. Cell adhesion to a 30-kPa Ecad-Fc PA gel required Cdc42- and formin-dependent filopodia formation, whereas adhesion to a 60-kPa Ecad-Fc PA gel induced Arp2/3-dependent lamellipodial protrusions. A quantitative 3D cell–cell adhesion assay and live cell imaging of cell–cell contact formation revealed that inhibition of Cdc42, formin, and Arp2/3 activities blocked the initiation, but not the maintenance of established cell–cell adhesions. These results indicate that the same signaling molecules activated by E-cadherin rigidity sensing on PA gels contribute to actin organization and membrane dynamics during cell–cell adhesion. We hypothesize that a transition in the stiffness of E-cadherin homotypic interactions regulates actin and membrane dynamics during initial stages of cell–cell adhesion.

scholarly journals RNAi-Mediated Knock-Down of Arylamine N-acetyltransferase-1 Expression Induces E-cadherin Up-Regulation and Cell-Cell Contact Growth Inhibition

PLoS ONE ◽  
2011 ◽  
Vol 6 (2) ◽  
pp. e17031 ◽  
Author(s):  
Jacky M. Tiang ◽  
Neville J. Butcher ◽  
Carleen Cullinane ◽  
Patrick O. Humbert ◽  
Rodney F. Minchin
Keyword(s):  
Growth Inhibition ◽  
Cell Contact ◽  
Knock Down ◽  
E Cadherin ◽  
Cell Cell

scholarly journals Minus end–directed motor KIFC3 suppresses E-cadherin degradation by recruiting USP47 to adherens junctions

2014 ◽  
Vol 25 (24) ◽  
pp. 3851-3860 ◽  
Author(s):  
Kyoko Sako-Kubota ◽  
Nobutoshi Tanaka ◽  
Shigenori Nagae ◽  
Wenxiang Meng ◽  
Masatoshi Takeichi
Keyword(s):  
Cell Adhesion ◽  
Proteasome Inhibitors ◽  
Adherens Junction ◽  
Ubiquitin Protein Ligase ◽  
Juxtamembrane Region ◽  
Specific Protease ◽  
Ubiquitin Specific Protease ◽  
Epithelial Sheets ◽  
E Cadherin ◽  
Cell Cell

The adherens junction (AJ) plays a crucial role in maintaining cell–cell adhesion in epithelial tissues. Previous studies show that KIFC3, a minus end–directed kinesin motor, moves into AJs via microtubules that grow from clusters of CAMSAP3 (also known as Nezha), a protein that binds microtubule minus ends. The function of junction-associated KIFC3, however, remains to be elucidated. Here we find that KIFC3 binds the ubiquitin-specific protease USP47, a protease that removes ubiquitin chains from substrates and hence inhibits proteasome-mediated proteolysis, and recruits it to AJs. Depletion of KIFC3 or USP47 promotes cleavage of E-cadherin at a juxtamembrane region of the cytoplasmic domain, resulting in the production of a 90-kDa fragment and the internalization of E-cadherin. This cleavage depends on the E3 ubiquitin protein ligase Hakai and is inhibited by proteasome inhibitors. E-cadherin ubiquitination consistently increases after depletion of KIFC3 or USP47. These findings suggest that KIFC3 suppresses the ubiquitination and resultant degradation of E-cadherin by recruiting USP47 to AJs, a process that may be involved in maintaining stable cell–cell adhesion in epithelial sheets.

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