Dimer asymmetry defines α-catenin interactions

The F-actin–binding cytoskeletal protein α-catenin interacts with β-catenin–cadherin complexes and stabilizes cell-cell junctions. The β-catenin–α-catenin complex cannot bind F-actin, whereas interactions of α-catenin with the cytoskeletal protein vinculin appear to be necessary to stabilize adherens junctions. Here we report the crystal structure of nearly full-length human α-catenin at 3.7-Å resolution. α-catenin forms an asymmetric dimer where the four-helix bundle domains of each subunit engage in distinct intermolecular interactions. This results in a left handshake–like dimer, wherein the two subunits have remarkably different conformations. The crystal structure explains why dimeric α-catenin has a higher affinity for F-actin than does monomeric α-catenin, why the β-catenin–α-catenin complex does not bind F-actin, how activated vinculin links the cadherin–catenin complex to the cytoskeleton and why α-catenin but not inactive vinculin can bind F-actin.

[1]  Eric Blanc,et al.  Automated structure solution with autoSHARP. , 2007, Methods in molecular biology.

[2]  S. Craig,et al.  F-actin binding site masked by the intramolecular association of vinculin head and tail domains , 1995, Nature.

[3]  S. Craig,et al.  The carboxy-terminal tail domain of vinculin contains a cryptic binding site for acidic phospholipids. , 1995, Biochemical and biophysical research communications.

[4]  M. Takeichi,et al.  The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. , 1988, Development.

[5]  Vincent B. Chen,et al.  Correspondence e-mail: , 2000 .

[6]  M. Itoh,et al.  Involvement of ZO-1 in Cadherin-based Cell Adhesion through Its Direct Binding to α Catenin and Actin Filaments , 1997, The Journal of cell biology.

[7]  M. Takeichi,et al.  Induction of polarized cell-cell association and retardation of growth by activation of the E-cadherin-catenin adhesion system in a dispersed carcinoma line , 1994, The Journal of cell biology.

[8]  P. Rørth,et al.  Regulatory mechanisms required for DE-cadherin function in cell migration and other types of adhesion , 2005, The Journal of cell biology.

[9]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[10]  E. Rangarajan,et al.  alpha-Catenin unfurls upon binding to vinculin , 2012 .

[11]  M. Takeichi,et al.  Remodeling of the adherens junctions during morphogenesis. , 2009, Current topics in developmental biology.

[12]  R. Liddington,et al.  αE-catenin is an autoinhibited molecule that coactivates vinculin , 2012, Proceedings of the National Academy of Sciences.

[13]  Clemens Vonrhein,et al.  Data processing and analysis with the autoPROC toolbox , 2011, Acta crystallographica. Section D, Biological crystallography.

[14]  B. Geiger,et al.  A 135‐kd membrane protein of intercellular adherens junctions. , 1984, The EMBO journal.

[15]  W. Weis,et al.  Re-solving the Cadherin-Catenin-Actin Conundrum* , 2006, Journal of Biological Chemistry.

[16]  E. Fuchs,et al.  Hyperproliferation and Defects in Epithelial Polarity upon Conditional Ablation of α-Catenin in Skin , 2001, Cell.

[17]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[18]  S. Yonemura,et al.  α-Catenin as a tension transducer that induces adherens junction development , 2010, Nature Cell Biology.

[19]  N. Adey,et al.  Polyphosphoinositides Inhibit the Interaction of Vinculin with Actin Filaments* , 1999, The Journal of Biological Chemistry.

[20]  William I. Weis,et al.  α-Catenin Is a Molecular Switch that Binds E-Cadherin-β-Catenin and Regulates Actin-Filament Assembly , 2005, Cell.

[21]  William I. Weis,et al.  Deconstructing the Cadherin-Catenin-Actin Complex , 2005, Cell.

[22]  A. Nose,et al.  Calcium-dependent cell-cell adhesion molecules (cadherins): subclass specificities and possible involvement of actin bundles , 1987, The Journal of cell biology.

[23]  A. P. Soler,et al.  Interaction of alpha-actinin with the cadherin/catenin cell-cell adhesion complex via alpha-catenin , 1995, The Journal of cell biology.

[24]  D L Rimm,et al.  α-Catenin Binds Directly to Spectrin and Facilitates Spectrin-Membrane Assembly in Vivo * , 2001, The Journal of Biological Chemistry.

[25]  M. Itoh,et al.  Functional Domains of α-Catenin Required for the Strong State of Cadherin-based Cell Adhesion , 1999, The Journal of cell biology.

[26]  B. Jockusch,et al.  Interaction of alpha-actinin and vinculin with actin: opposite effects on filament network formation. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[27]  K. Tachibana,et al.  Two Cell Adhesion Molecules, Nectin and Cadherin, Interact through Their Cytoplasmic Domain–Associated Proteins , 2000, The Journal of cell biology.

[28]  N. Kioka,et al.  Spatial distribution and functional significance of activated vinculin in living cells , 2005, The Journal of cell biology.

[29]  Jie J. Zheng,et al.  Crystal structure of a full-length beta-catenin. , 2008, Structure.

[30]  D. Barford,et al.  Crystal structure of the M‐fragment of α‐catenin: implications for modulation of cell adhesion , 2001, The EMBO journal.

[31]  Gerard Bricogne,et al.  Crystal structure of human vinculin. , 2004, Structure.

[32]  E. Rangarajan,et al.  A Helix Replacement Mechanism Directs Metavinculin Functions , 2010, PloS one.

[33]  M. Itoh,et al.  Functional domains of alpha-catenin required for the strong state of cadherin-based cell adhesion. , 1999 .

[34]  T. Lecuit α-catenin mechanosensing for adherens junctions , 2010, Nature Cell Biology.

[35]  S. Ishihara,et al.  The roles of catenins in the cadherin-mediated cell adhesion: functional analysis of E-cadherin-alpha catenin fusion molecules , 1994, The Journal of cell biology.

[36]  Wolfgang Kabsch,et al.  Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants , 1993 .

[37]  W. Birchmeier,et al.  E-cadherin and APC compete for the interaction with beta-catenin and the cytoskeleton , 1994, The Journal of cell biology.

[38]  D. Rimm,et al.  Vinculin Is Associated with the E-cadherin Adhesion Complex* , 1997, The Journal of Biological Chemistry.

[39]  P. Evans,et al.  Scaling and assessment of data quality. , 2006, Acta crystallographica. Section D, Biological crystallography.

[40]  Peter D. Kwong,et al.  Structural basis of cell-cell adhesion by cadherins , 1995, Nature.

[41]  Akihiro Kusumi,et al.  Cytoplasmic Regulation of the Movement of E-Cadherin on the Free Cell Surface as Studied by Optical Tweezers and Single Particle Tracking: Corralling and Tethering by the Membrane Skeleton , 1998, The Journal of cell biology.

[42]  M. Takeichi,et al.  EPLIN mediates linkage of the cadherin–catenin complex to F-actin and stabilizes the circumferential actin belt , 2008, Proceedings of the National Academy of Sciences.

[43]  K. DeMali,et al.  Vinculin regulates cell-surface E-cadherin expression by binding to β-catenin , 2010, Journal of Cell Science.

[44]  W. Weis,et al.  Structure and mechanism of cadherins and catenins in cell-cell contacts. , 2007, Annual review of cell and developmental biology.

[45]  B. Jockusch,et al.  Vinculin Is Part of the Cadherin–Catenin Junctional Complex: Complex Formation between α-Catenin and Vinculin , 1998, The Journal of cell biology.

[46]  D L Rimm,et al.  Alpha 1(E)-catenin is an actin-binding and -bundling protein mediating the attachment of F-actin to the membrane adhesion complex. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[47]  N. Inoue,et al.  Actomyosin tension is required for correct recruitment of adherens junction components and zonula occludens formation. , 2006, Experimental cell research.

[48]  E. Fuchs,et al.  α-catenin: at the junction of intercellular adhesion and actin dynamics , 2004, Nature Reviews Molecular Cell Biology.

[49]  B. Honig,et al.  Thinking outside the cell: how cadherins drive adhesion. , 2012, Trends in cell biology.

[50]  S. Butz,et al.  An alpha-E-catenin gene trap mutation defines its function in preimplantation development. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[51]  William I. Weis,et al.  Structure of the Dimerization and β-Catenin- Binding Region of α-Catenin , 2000 .

[52]  Shin Lin,et al.  High-affinity interaction of vinculin with actin filaments in vitro , 1982, Cell.

[53]  E. Rangarajan,et al.  The Cytoskeletal Protein α-Catenin Unfurls upon Binding to Vinculin* , 2012, The Journal of Biological Chemistry.

[54]  W. Weis,et al.  Biochemical and Structural Definition of the l-Afadin- and Actin-binding Sites of α-Catenin* , 2002, The Journal of Biological Chemistry.

[55]  GERHARDISENBERGt Interaction ofa-actinin andvinculin withactin: Opposite effects on filament network formation , 1981 .

[56]  David L Rimm,et al.  Direct Interaction of the C-Terminal Domain of α-Catenin and F-Actin is Necessary for Stabilized Cell-Cell Adhesion , 2006, Cell communication & adhesion.

[57]  S. Hirohashi,et al.  Identification of a neural α-catenin as a key regulator of cadherin function and multicellular organization , 1992, Cell.

[58]  S. Barry,et al.  Acidic phospholipids inhibit the intramolecular association between the N- and C-terminal regions of vinculin, exposing actin-binding and protein kinase C phosphorylation sites. , 1996, The Biochemical journal.

[59]  S. Craig,et al.  α-Catenin Uses a Novel Mechanism to Activate Vinculin* , 2012, The Journal of Biological Chemistry.

[60]  M. Takeichi Morphogenetic roles of classic cadherins. , 1995, Current opinion in cell biology.

[61]  T. Uemura,et al.  α-Catenin-Vinculin Interaction Functions to Organize the Apical Junctional Complex in Epithelial Cells , 1998, The Journal of cell biology.