Three-dimensional structure of vinculin bound to actin filaments.

Vinculin plays a pivotal role in cell adhesion and migration by providing the link between the actin cytoskeleton and the transmembrane receptors, integrin and cadherin. We used a combination of electron microscopy, computational docking, and biochemistry to provide an atomic model of how the vinculin tail binds actin filaments. The vinculin tail actin binding site comprises two distinct regions. One of these regions is exposed in the full-length autoinhibited conformation of vinculin, whereas the second site is sterically occluded by vinculin's N-terminal domain. The partial accessibility of the F-actin binding site in the autoinhibited full-length vinculin structure suggests that F-actin can act as part of a combinatorial input framework with other binding partners such as alpha-catenin or talin to induce vinculin head-tail dissociation, thus promoting vinculin activation. Furthermore, binding to F-actin potentiates a local rearrangement in the vinculin tail that in turn promotes vinculin dimerization and, hence, formation of actin bundles.

[1]  D. DeRosier,et al.  How to analyze electron micrographs of rafts of actin filaments crosslinked by actin-binding proteins. , 1998, Journal of molecular biology.

[2]  B. Geiger,et al.  Molecular interactions in the submembrane plaque of cell-cell and cell-matrix adhesions. , 1995, Acta anatomica.

[3]  E. Egelman A robust algorithm for the reconstruction of helical filaments using single-particle methods. , 2000, Ultramicroscopy.

[4]  V. Orlov,et al.  Differential scanning calorimetric studies on myosin and actin. , 1998, Biochemistry. Biokhimiia.

[5]  J R Kremer,et al.  Computer visualization of three-dimensional image data using IMOD. , 1996, Journal of structural biology.

[6]  K A Taylor,et al.  Formation of 2-D paracrystals of F-actin on phospholipid layers mixed with quaternary ammonium surfactants. , 1992, Journal of structural biology.

[7]  F. Schachat,et al.  Isoforms of �-actinin from cardiac, smooth, and skeletal muscle form polar arrays of actin filaments , 2000 .

[8]  P. Sansonetti,et al.  Binding of the Shigella protein IpaA to vinculin induces F‐actin depolymerization , 1999, The EMBO journal.

[9]  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.

[10]  Niels Volkmann,et al.  A novel three-dimensional variant of the watershed transform for segmentation of electron density maps. , 2002, Journal of structural biology.

[11]  D. DeRosier,et al.  Determination of the alpha-actinin-binding site on actin filaments by cryoelectron microscopy and image analysis , 1994, The Journal of cell biology.

[12]  Niels Volkmann,et al.  A protein disulfide oxidoreductase from the archaeon Pyrococcus furiosus contains two thioredoxin fold units , 1998, Nature Structural Biology.

[13]  B. Jockusch,et al.  The ultrastructure of chicken gizzard vinculin as visualized by high-resolution electron microscopy. , 1996, Journal of structural biology.

[14]  Niels Volkmann,et al.  An Atomic Model of Actin Filaments Cross-Linked by Fimbrin and Its Implications for Bundle Assembly and Function , 2001, The Journal of cell biology.

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

[16]  D G Morgan,et al.  Image analysis of helical objects: the Brandeis Helical Package. , 1996, Journal of structural biology.

[17]  S. Craig,et al.  Actin Activates a Cryptic Dimerization Potential of the Vinculin Tail Domain* , 2000, The Journal of Biological Chemistry.

[18]  T. Tsong,et al.  Enthalpic and entropic contributions to actin stability: calorimetry, circular dichroism, and fluorescence study and effects of calcium. , 1990, Biochemistry.

[19]  Roberto Dominguez,et al.  Actin-binding proteins--a unifying hypothesis. , 2004, Trends in biochemical sciences.

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

[21]  S. Jones,et al.  Protein domain interfaces: characterization and comparison with oligomeric protein interfaces. , 2000, Protein engineering.

[22]  B. Lowes,et al.  Dilated cardiomyopathy associated with deficiency of the cytoskeletal protein metavinculin. , 1996, Circulation.

[23]  B. Jockusch,et al.  Crosstalk between cell adhesion molecules: vinculin as a paradigm for regulation by conformation. , 1996, Trends in cell biology.

[24]  C. Gicquaud,et al.  Stabilization of actin by phalloidin: a differential scanning calorimetric study. , 1991, Biochemical and biophysical research communications.

[25]  Niels Volkmann,et al.  Docking of atomic models into reconstructions from electron microscopy. , 2003, Methods in enzymology.

[26]  Clive R. Bagshaw,et al.  Crystal Structure of the Vinculin Tail Suggests a Pathway for Activation , 1999, Cell.

[27]  N. Volkmann,et al.  The Structural Basis of Myosin V Processive Movement as Revealed by Electron Cryomicroscopy , 2022 .

[28]  M. Keating,et al.  Metavinculin Mutations Alter Actin Interaction in Dilated Cardiomyopathy , 2002, Circulation.

[29]  Keith Burridge,et al.  Recruitment of the Arp2/3 complex to vinculin , 2002, The Journal of cell biology.

[30]  S. Craig,et al.  Two Distinct Head-Tail Interfaces Cooperate to Suppress Activation of Vinculin by Talin* , 2005, Journal of Biological Chemistry.

[31]  N. Volkmann,et al.  Quantitative fitting of atomic models into observed densities derived by electron microscopy. , 1999, Journal of structural biology.

[32]  N. Dalton,et al.  Animal Model Heterozygous Inactivation of the Vinculin Gene Predisposes to Stress-Induced Cardiomyopathy , 2004 .

[33]  Constantina Bakolitsa,et al.  Structural basis for vinculin activation at sites of cell adhesion , 2004, Nature.

[34]  P. Mangeat,et al.  An interaction between vinculin and talin , 1984, Nature.

[35]  R. Waterston,et al.  Vinculin is essential for muscle function in the nematode , 1991, The Journal of cell biology.

[36]  E. Adamson,et al.  Vinculin knockout results in heart and brain defects during embryonic development. , 1998, Development.

[37]  E. Zamir,et al.  Molecular complexity and dynamics of cell-matrix adhesions. , 2001, Journal of cell science.

[38]  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.

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

[40]  B. Jockusch,et al.  Characterization of two F-actin-binding and oligomerization sites in the cell-contact protein vinculin. , 1997, European journal of biochemistry.

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

[42]  W Chiu,et al.  EMAN: semiautomated software for high-resolution single-particle reconstructions. , 1999, Journal of structural biology.

[43]  R A Milligan,et al.  Protein-protein interactions in the rigor actomyosin complex. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Niels Volkmann,et al.  Evidence for cleft closure in actomyosin upon ADP release , 2000, Nature Structural Biology.