CD2AP Links Cortactin and Capping Protein at the Cell Periphery To Facilitate Formation of Lamellipodia

ABSTRACT Understanding the physiology of complex relationships between components of signaling pathways and the actin cytoskeleton is an important challenge. CD2AP is a membrane scaffold protein implicated in a variety of physiological and disease processes. The physiological function of CD2AP is unclear, but its biochemical interactions suggest that it has a role in dynamic actin assembly. Here, we report that CD2AP functions to facilitate the recruitment of actin capping protein (CP) to the Src kinase substrate, cortactin, at the cell periphery, and that this is necessary for formation of the short branched filaments that characterize lamellipodium formation and are required for cell migration. Superresolution fluorescence microscopy demonstrated that the efficient colocalization of CP and cortactin at the cell periphery required CD2AP. As both cortactin and CP function to enhance branched actin filament formation, CD2AP functions synergistically to enhance the function of both proteins. Our data demonstrate how the interplay between specialized actin regulatory molecules shapes the actin cytoskeleton.

[1]  Shawn M. Gomez,et al.  Arp2/3 Is Critical for Lamellipodia and Response to Extracellular Matrix Cues but Is Dispensable for Chemotaxis , 2012, Cell.

[2]  Suliana Manley,et al.  Functional nanoscale organization of signaling molecules downstream of the T cell antigen receptor. , 2011, Immunity.

[3]  Prabuddha Sengupta,et al.  Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis , 2011, Nature Methods.

[4]  D. G. Clark,et al.  Common variants in MS4A4/MS4A6E, CD2uAP, CD33, and EPHA1 are associated with late-onset Alzheimer’s disease , 2011, Nature Genetics.

[5]  Nick C Fox,et al.  Common variants in ABCA7, MS4A6A/MS4A4E, EPHA1, CD33 and CD2AP are associated with Alzheimer’s disease , 2011, Nature Genetics.

[6]  X. Zhuang,et al.  Superresolution Imaging of Chemical Synapses in the Brain , 2010, Neuron.

[7]  Qin Su,et al.  A Multifunctional Lentiviral-Based Gene Knockdown with Concurrent Rescue that Controls for Off-Target Effects of RNAi , 2010, Genom. Proteom. Bioinform..

[8]  J. Cooper,et al.  Structural characterization of a capping protein interaction motif defines a family of actin filament regulators , 2010, Nature Structural &Molecular Biology.

[9]  Mark Bates,et al.  Super-resolution fluorescence microscopy. , 2009, Annual review of biochemistry.

[10]  S. Weed,et al.  Cortactin branches out: roles in regulating protrusive actin dynamics. , 2008, Cell motility and the cytoskeleton.

[11]  J. Small,et al.  Unravelling the structure of the lamellipodium , 2008, Journal of microscopy.

[12]  R. Mullins,et al.  Capping Protein Increases the Rate of Actin-Based Motility by Promoting Filament Nucleation by the Arp2/3 Complex , 2008, Cell.

[13]  S. Hell.  Microscopy and its focal switch , 2008, Nature Methods.

[14]  Mark Bates,et al.  Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes , 2007, Science.

[15]  Frank B Gertler,et al.  Ena/VASP proteins have an anti-capping independent function in filopodia formation. , 2007, Molecular biology of the cell.

[16]  Eric A. Vitriol,et al.  Enhanced EGFP-chromophore-assisted laser inactivation using deficient cells rescued with functional EGFP-fusion proteins , 2007, Proceedings of the National Academy of Sciences.

[17]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[18]  Tony Pawson,et al.  Identification of a Novel Inhibitory Actin-capping Protein Binding Motif in CD2-associated Protein* , 2006, Journal of Biological Chemistry.

[19]  J. Cooper,et al.  Cortactin has an essential and specific role in osteoclast actin assembly. , 2006, Molecular biology of the cell.

[20]  A. Shaw,et al.  CD2-associated Protein (CD2AP) Expression in Podocytes Rescues Lethality of CD2AP Deficiency* , 2005, Journal of Biological Chemistry.

[21]  D. Hanein,et al.  Ena/VASP Proteins Enhance Actin Polymerization in the Presence of Barbed End Capping Proteins*[boxs] , 2005, Journal of Biological Chemistry.

[22]  Nicole S. Bryce,et al.  Cortactin Promotes Cell Motility by Enhancing Lamellipodial Persistence , 2005, Current Biology.

[23]  Gary G. Borisy,et al.  Lamellipodial Versus Filopodial Mode of the Actin Nanomachinery Pivotal Role of the Filament Barbed End , 2004, Cell.

[24]  J. Cooper,et al.  How Capping Protein Binds the Barbed End of the Actin Filament , 2003, Current Biology.

[25]  Andrew W. Kinley,et al.  Fgd1, the Cdc42 GEF responsible for Faciogenital Dysplasia, directly interacts with cortactin and mAbp1 to modulate cell shape. , 2003, Human molecular genetics.

[26]  Nicholas J. Hutchings,et al.  Linking the T Cell Surface Protein CD2 to the Actin-capping Protein CAPZ via CMS and CIN85* , 2003, Journal of Biological Chemistry.

[27]  G. Corthals,et al.  A Cortactin-CD2-associated Protein (CD2AP) Complex Provides a Novel Link between Epidermal Growth Factor Receptor Endocytosis and the Actin Cytoskeleton* , 2003, Journal of Biological Chemistry.

[28]  Alissa M. Weaver,et al.  Cortactin Interacts with WIP in Regulating Arp2/3 Activation and Membrane Protrusion , 2003, Current Biology.

[29]  A. Karginov,et al.  Dynamin2 and Cortactin Regulate Actin Assembly and Filament Organization , 2002, Current Biology.

[30]  Alissa M. Weaver,et al.  Interaction of Cortactin and N-WASp with Arp2/3 Complex , 2002, Current Biology.

[31]  Gary G. Borisy,et al.  Antagonism between Ena/VASP Proteins and Actin Filament Capping Regulates Fibroblast Motility , 2002, Cell.

[32]  Klemens Rottner,et al.  The lamellipodium: where motility begins. , 2002, Trends in cell biology.

[33]  R. Cotran,et al.  CD2AP localizes to the slit diaphragm and binds to nephrin via a novel C-terminal domain. , 2001, The American journal of pathology.

[34]  S. Weed,et al.  Cortactin: coupling membrane dynamics to cortical actin assembly , 2001, Oncogene.

[35]  Peijun Zhang,et al.  Activation of Arp2/3 complex-mediated actin polymerization by cortactin , 2001, Nature Cell Biology.

[36]  Alissa M. Weaver,et al.  Cortactin Localization to Sites of Actin Assembly in Lamellipodia Requires Interactions with F-Actin and the Arp2/3 Complex , 2000, The Journal of cell biology.

[37]  A. Shaw,et al.  CD2AP is expressed with nephrin in developing podocytes and is found widely in mature kidney and elsewhere. , 2000, American journal of physiology. Renal physiology.

[38]  T. Kitamura,et al.  Plat-E: an efficient and stable system for transient packaging of retroviruses , 2000, Gene Therapy.

[39]  Michael Loran Dustin,et al.  Congenital nephrotic syndrome in mice lacking CD2-associated protein. , 1999, Science.

[40]  Marie-France Carlier,et al.  Reconstitution of actin-based motility of Listeria and Shigella using pure proteins , 1999, Nature.

[41]  A. Mui,et al.  STAT5 as a molecular regulator of proliferation, differentiation and apoptosis in hematopoietic cells , 1999, The EMBO journal.

[42]  S. Weed,et al.  Identification of a Novel Cortactin SH3 Domain-Binding Protein and Its Localization to Growth Cones of Cultured Neurons , 1998, Molecular and Cellular Biology.

[43]  Patricia L. Widder,et al.  A Novel Adaptor Protein Orchestrates Receptor Patterning and Cytoskeletal Polarity in T-Cell Contacts , 1998, Cell.

[44]  Jonathan A. Cooper,et al.  Dynamics of capping protein and actin assembly in vitro: uncapping barbed ends by polyphosphoinositides , 1996, The Journal of cell biology.

[45]  J Strackee,et al.  Largest contour segmentation: a tool for the localization of spots in confocal images. , 1996, Cytometry.

[46]  J. Small.  Lamellipodia architecture: actin filament turnover and the lateral flow of actin filaments during motility. , 1994, Seminars in cell biology.

[47]  J. Parsons,et al.  Cortactin, an 80/85-kilodalton pp60src substrate, is a filamentous actin-binding protein enriched in the cell cortex , 1993, The Journal of cell biology.

[48]  T. Pollard,et al.  Effect of capping protein on the kinetics of actin polymerization. , 1985, Biochemistry.