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PMID: 18923426 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Clustering of VASP actively drives processive, WH2 domain-mediated actin filament elongation.

The EMBO journal ·Vol. 27 ·No. 22 ·2008-11-19 ·Pages 2943-54

Breitsprecher D, Kiesewetter AK, Linkner J, Urbanke C, Resch GP, Small JV, Faix J

Abstract

Vasodilator-stimulated phosphoprotein (VASP) is a key regulator of dynamic actin structures like filopodia and lamellipodia, but its precise function in their formation is controversial. Using in vitro TIRF microscopy, we show for the first time that both human and Dictyostelium VASP are directly involved in accelerating filament elongation by delivering monomeric actin to the growing barbed end. In solution, DdVASP markedly accelerated actin filament elongation in a concentration-dependent manner but was inhibited by low concentrations of capping protein (CP). In striking contrast, VASP clustered on functionalized beads switched to processive filament elongation that became insensitive even to very high concentrations of CP. Supplemented with the in vivo analysis of VASP mutants and an EM structure of the protein, we propose a mechanism by which membrane-associated VASP oligomers use their WH2 domains to effect both the tethering of actin filaments and their processive elongation in sites of active actin assembly.

MeSH Terms
Actin Capping Proteins/metabolism Actin Cytoskeleton/chemistry,metabolism Actins/metabolism Animals Cell Adhesion Molecules/chemistry,genetics,metabolism Dictyostelium/cytology,metabolism Humans Microfilament Proteins/chemistry,genetics,metabolism Microscopy, Fluorescence/methods Models, Biological Mutation Phosphoproteins/chemistry,genetics,metabolism Profilins/metabolism Protein Structure, Tertiary Protozoan Proteins/metabolism
Chemicals
Actin Capping Proteins Actins Cell Adhesion Molecules Microfilament Proteins Phosphoproteins Profilins Protozoan Proteins vasodilator-stimulated phosphoprotein
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Breitsprecher Dennis
Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany.
Kiesewetter Antje K
Linkner Joern
Urbanke Claus
Resch Guenter P
Small J Victor
Faix Jan
References (50)
50 references, click to expand
  1. Force generation by actin polymerization II: the elastic ratchet and tethered filaments.
    Biophys J. 2003 Mar;84(3):1591-605 PMID: 12609863
  2. The EVH2 domain of the vasodilator-stimulated phosphoprotein mediates tetramerization, F-actin binding, and actin bundle formation.
    J Biol Chem. 1999 Aug 13;274(33):23549-57 PMID: 10438535
  3. How VASP enhances actin-based motility.
    J Cell Biol. 2003 Oct 13;163(1):131-42 PMID: 14557252
  4. The beta-thymosin/WH2 domain; structural basis for the switch from inhibition to promotion of actin assembly.
    Cell. 2004 May 28;117(5):611-23 PMID: 15163409
  5. Lamellipodin, an Ena/VASP ligand, is implicated in the regulation of lamellipodial dynamics.
    Dev Cell. 2004 Oct;7(4):571-83 PMID: 15469845
  6. Reconstitution of actin-based motility of Listeria and Shigella using pure proteins.
    Nature. 1999 Oct 7;401(6753):613-6 PMID: 10524632
  7. PREL1 provides a link from Ras signalling to the actin cytoskeleton via Ena/VASP proteins.
    FEBS Lett. 2005 Jan 17;579(2):455-63 PMID: 15642358
  8. Real-time measurements of actin filament polymerization by total internal reflection fluorescence microscopy.
    Biophys J. 2005 Feb;88(2):1387-402 PMID: 15556992
  9. Bacteria-host-cell interactions at the plasma membrane: stories on actin cytoskeleton subversion.
    Dev Cell. 2005 Jul;9(1):3-17 PMID: 15992537
  10. Ena/VASP proteins enhance actin polymerization in the presence of barbed end capping proteins.
    J Biol Chem. 2005 Aug 5;280(31):28653-62 PMID: 15939738
  11. VASP dynamics during lamellipodia protrusion.
    Nat Cell Biol. 1999 Sep;1(5):321-2 PMID: 10559946
  12. Dual epitope recognition by the VASP EVH1 domain modulates polyproline ligand specificity and binding affinity.
    EMBO J. 2000 Sep 15;19(18):4903-14 PMID: 10990454
  13. cAMP-dependent protein kinase phosphorylation of EVL, a Mena/VASP relative, regulates its interaction with actin and SH3 domains.
    J Biol Chem. 2000 Nov 17;275(46):36143-51 PMID: 10945997
  14. Clamped-filament elongation model for actin-based motors.
    Biophys J. 2002 Feb;82(2):605-17 PMID: 11806905
  15. WH2 domain: a small, versatile adapter for actin monomers.
    FEBS Lett. 2002 Feb 20;513(1):92-7 PMID: 11911886
  16. Antagonism between Ena/VASP proteins and actin filament capping regulates fibroblast motility.
    Cell. 2002 May 17;109(4):509-21 PMID: 12086607
  17. Contribution of Ena/VASP proteins to intracellular motility of listeria requires phosphorylation and proline-rich core but not F-actin binding or multimerization.
    Mol Biol Cell. 2002 Jul;13(7):2383-96 PMID: 12134077
  18. Critical roles of phosphorylation and actin binding motifs, but not the central proline-rich region, for Ena/vasodilator-stimulated phosphoprotein (VASP) function during cell migration.
    Mol Biol Cell. 2002 Jul;13(7):2533-46 PMID: 12134088
  19. Requirement of a vasodilator-stimulated phosphoprotein family member for cell adhesion, the formation of filopodia, and chemotaxis in dictyostelium.
    J Biol Chem. 2002 Dec 20;277(51):49877-87 PMID: 12388544
  20. Mechanism of filopodia initiation by reorganization of a dendritic network.
    J Cell Biol. 2003 Feb 3;160(3):409-21 PMID: 12566431
  21. Cellular motility driven by assembly and disassembly of actin filaments.
    Cell. 2003 Feb 21;112(4):453-65 PMID: 12600310
  22. Control of the assembly of ATP- and ADP-actin by formins and profilin.
    Cell. 2006 Jan 27;124(2):423-35 PMID: 16439214
  23. The bundling activity of vasodilator-stimulated phosphoprotein is required for filopodium formation.
    Proc Natl Acad Sci U S A. 2006 May 16;103(20):7694-9 PMID: 16675552
  24. Understanding the role of the G-actin-binding domain of Ena/VASP in actin assembly.
    J Struct Biol. 2006 Aug;155(2):195-201 PMID: 16684607
  25. Mechanism of actin network attachment to moving membranes: barbed end capture by N-WASP WH2 domains.
    Cell. 2007 Mar 9;128(5):901-13 PMID: 17350575
  26. Regulation of actin filament assembly by Arp2/3 complex and formins.
    Annu Rev Biophys Biomol Struct. 2007;36:451-77 PMID: 17477841
  27. Ena/VASP proteins have an anti-capping independent function in filopodia formation.
    Mol Biol Cell. 2007 Jul;18(7):2579-91 PMID: 17475772
  28. Control of actin assembly dynamics in cell motility.
    J Biol Chem. 2007 Aug 10;282(32):23005-9 PMID: 17576764
  29. The beta-thymosin/WH2 fold: multifunctionality and structure.
    Ann N Y Acad Sci. 2007 Sep;1112:86-94 PMID: 17468236
  30. Structural basis for the recruitment of profilin-actin complexes during filament elongation by Ena/VASP.
    EMBO J. 2007 Oct 31;26(21):4597-606 PMID: 17914456
  31. Ena/VASP Is Required for neuritogenesis in the developing cortex.
    Neuron. 2007 Nov 8;56(3):441-55 PMID: 17988629
  32. Analysis of the function of Spire in actin assembly and its synergy with formin and profilin.
    Mol Cell. 2007 Nov 30;28(4):555-68 PMID: 18042452
  33. High-resolution structural analysis of mammalian profilin 2a complex formation with two physiological ligands: the formin homology 1 domain of mDia1 and the proline-rich domain of VASP.
    J Mol Biol. 2008 Jan 4;375(1):270-90 PMID: 18001770
  34. Filopodia are required for cortical neurite initiation.
    Nat Cell Biol. 2007 Dec;9(12):1347-59 PMID: 18026093
  35. Differentially oriented populations of actin filaments generated in lamellipodia collaborate in pushing and pausing at the cell front.
    Nat Cell Biol. 2008 Mar;10(3):306-13 PMID: 18278037
  36. Ena/VASP proteins capture actin filament barbed ends.
    J Biol Chem. 2008 Apr 11;283(15):9814-9 PMID: 18283104
  37. Relaxing the actin cytoskeleton for adhesion and movement with Ena/VASP.
    J Cell Biol. 2008 Apr 7;181(1):19-25 PMID: 18378777
  38. Arp2/3 complex interactions and actin network turnover in lamellipodia.
    EMBO J. 2008 Apr 9;27(7):982-92 PMID: 18309290
  39. RIAM, an Ena/VASP and Profilin ligand, interacts with Rap1-GTP and mediates Rap1-induced adhesion.
    Dev Cell. 2004 Oct;7(4):585-95 PMID: 15469846
  40. Insertional assembly of actin filament barbed ends in association with formins produces piconewton forces.
    Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14725-30 PMID: 15377785
  41. Formin is a processive motor that requires profilin to accelerate actin assembly and associated ATP hydrolysis.
    Cell. 2004 Oct 29;119(3):419-29 PMID: 15507212
  42. The 46/50 kDa phosphoprotein VASP purified from human platelets is a novel protein associated with actin filaments and focal contacts.
    EMBO J. 1992 Jun;11(6):2063-70 PMID: 1318192
  43. The proline-rich focal adhesion and microfilament protein VASP is a ligand for profilins.
    EMBO J. 1995 Apr 18;14(8):1583-9 PMID: 7737110
  44. Identification, purification, and characterization of a zyxin-related protein that binds the focal adhesion and microfilament protein VASP (vasodilator-stimulated phosphoprotein).
    Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7956-60 PMID: 7644520
  45. The focal-adhesion vasodilator-stimulated phosphoprotein (VASP) binds to the proline-rich domain in vinculin.
    Biochem J. 1996 Sep 15;318 ( Pt 3):753-7 PMID: 8836115
  46. Mena, a relative of VASP and Drosophila Enabled, is implicated in the control of microfilament dynamics.
    Cell. 1996 Oct 18;87(2):227-39 PMID: 8861907
  47. p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin.
    EMBO J. 1997 Jun 2;16(11):3044-56 PMID: 9214622
  48. A novel proline-rich motif present in ActA of Listeria monocytogenes and cytoskeletal proteins is the ligand for the EVH1 domain, a protein module present in the Ena/VASP family.
    EMBO J. 1997 Sep 1;16(17):5433-44 PMID: 9312002
  49. Role of proteins of the Ena/VASP family in actin-based motility of Listeria monocytogenes.
    J Cell Biol. 1999 Mar 22;144(6):1245-58 PMID: 10087267
  50. Actin-based motility: from molecules to movement.
    Bioessays. 2003 Apr;25(4):336-45 PMID: 12655641
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2008.211
Published
2008-11-19
Epub
2008-00-16
Pages
2943-54
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2585163
Subset
IM
External Links
PubMed source DOI: 10.1038/emboj.2008.211 PMC Full Text View journal details
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