title: RIAM and vinculin binding to talin are mutually exclusive and regulate adhesion assembly and turnover.
abstract: Talin activates integrins, couples them to F-actin, and recruits vinculin to focal adhesions (FAs). Here, we report the structural characterization of the talin rod: 13 helical bundles (R1-R13) organized into a compact cluster of four-helix bundles (R2-R4) within a linear chain of five-helix bundles. Nine of the bundles contain vinculin-binding sites (VBS); R2R3 are atypical, with each containing two VBS. Talin R2R3 also binds synergistically to RIAM, a Rap1 effector involved in integrin activation. Biochemical and structural data show that vinculin and RIAM binding to R2R3 is mutually exclusive. Moreover, vinculin binding requires domain unfolding, whereas RIAM binds the folded R2R3 double domain. In cells, RIAM is enriched in nascent adhesions at the leading edge whereas vinculin is enriched in FAs. We propose a model in which RIAM binding to R2R3 initially recruits talin to membranes where it activates integrins. As talin engages F-actin, force exerted on R2R3 disrupts RIAM binding and exposes the VBS, which recruit vinculin to stabilize the complex.
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Benjamin T
authors: BT
authors: Goult
authors: School of Biological Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors: Thomas
authors: T
authors: Zacharchenko
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Neil
authors: N
authors: Bate
authors: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
authors: Ricky
authors: R
authors: Tsang
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Fiona
authors: F
authors: Hey
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Alexandre R
authors: AR
authors: Gingras
authors: School of Biological Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors: Paul R
authors: PR
authors: Elliott
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Gordon C K
authors: GCK
authors: Roberts
authors: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
authors: Christoph
authors: C
authors: Ballestrem
authors: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, United Kingdom. Electronic address: drc@le.ac.uk.
authors: David R
authors: DR
authors: Critchley
authors: School of Biological Sciences, University of Liverpool, Crown Street, Liverpool L69 7ZB, United Kingdom. Electronic address: i.barsukov@liverpool.ac.uk.
authors: Igor L
authors: IL
authors: Barsukov
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D018360:Crystallography, X-Ray; D022001:Focal Adhesions / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D016608:Talin / Q000737:chemistry* / Q000378:metabolism; D016596:Vinculin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D008565:Membrane Proteins; C521189:RIAM protein, mouse; C518832:TLN1 protein, human; D016608:Talin; C496077:Tln1 protein, mouse; D016596:Vinculin
references: Biomol NMR Assign. 2008 Jun;2(1):17-9
references: 19636914
references: Exp Cell Res. 2008 May 1;314(8):1839-49
references: 18342854
references: J Biol Chem. 2009 Feb 20;284(8):5119-27
references: 19098287
references: J Biol Chem. 2012 Apr 20;287(17):13799-812
references: 22351767
references: Nature. 2010 Nov 25;468(7323):580-4
references: 21107430
references: Eur J Cell Biol. 2010 Sep;89(9):661-73
references: 20605055
references: Trends Cell Biol. 2006 Sep;16(9):453-60
references: 16893648
references: Structure. 2005 Jan;13(1):65-74
references: 15642262
references: Trends Biochem Sci. 2011 Apr;36(4):191-8
references: 21216149
references: EMBO J. 2008 Jan 23;27(2):458-69
references: 18157087
references: Structure. 2010 Oct 13;18(10):1289-99
references: 20947018
references: J Biol Chem. 2008 Mar 7;283(10):6118-25
references: 18165225
references: Annu Rev Biophys. 2009;38:235-54
references: 19416068
references: EMBO J. 2004 Aug 4;23(15):2942-51
references: 15272303
references: Proc Natl Acad Sci U S A. 2011 Oct 11;108(41):17129-34
references: 21969587
references: Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):72-82
references: 16369096
references: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references: 15299926
references: Biochemistry. 2006 Feb 14;45(6):1805-17
references: 16460027
references: J Mol Biol. 1994 Jan 28;235(4):1278-90
references: 8308890
references: J Cell Sci. 1999 Jun;112 ( Pt 11):1655-69
references: 10318759
references: PLoS Comput Biol. 2008 Feb;4(2):e24
references: 18282082
references: J Cell Biol. 2011 Oct 31;195(3):499-513
references: 22042621
references: J Cell Sci. 2009 Jun 15;122(Pt 12):1947-53
references: 19494122
references: J Cell Biol. 2009 Nov 30;187(5):715-31
references: 19948488
references: J Cell Biol. 2010 Mar 22;188(6):877-90
references: 20308429
references: Nat Rev Mol Cell Biol. 2010 Apr;11(4):288-300
references: 20308986
references: EMBO J. 2009 Nov 18;28(22):3623-32
references: 19798053
references: Dev Cell. 2004 Oct;7(4):585-95
references: 15469846
references: Nature. 2004 Jan 8;427(6970):171-5
references: 14702644
references: J Biol Chem. 2005 Nov 4;280(44):37217-24
references: 16135522
references: J Cell Sci. 2009 Jan 15;122(Pt 2):187-98
references: 19118211
references: Biophys Rev. 2009 Jul;1(2):61-69
references: 19655048
references: FEBS Lett. 2010 Jun 3;584(11):2237-41
references: 20399778
references: Mol Cell. 2008 Jul 11;31(1):124-33
references: 18614051
references: PLoS One. 2012;7(4):e34461
references: 22496808
references: J Cell Biol. 2008 Jun 30;181(7):1211-22
references: 18573917
references: J Biol Chem. 2009 Mar 27;284(13):8866-76
references: 19176533
references: Curr Biol. 2006 Sep 19;16(18):1796-806
references: 16979556
references: J Cell Biol. 2012 Oct 15;199(2):317-30
references: 23045549
references: J Cell Biol. 2007 Dec 3;179(5):1043-57
references: 18056416
references: J Cell Sci. 1996 Nov;109 ( Pt 11):2715-26
references: 8937989
references: Eur J Cell Biol. 2006 Jun;85(6):487-500
references: 16584805
references: J Biol Chem. 2009 May 29;284(22):15097-106
references: 19297334
references: Science. 2009 Jan 30;323(5914):638-41
references: 19179532
references: J Cell Sci. 2010 May 1;123(Pt 9):1385-8
references: 20410370
references: PLoS Biol. 2011 Dec;9(12):e1001223
references: 22205879
references: J Biol Chem. 2010 Sep 17;285(38):29577-87
references: 20610383
references: EMBO J. 2010 Mar 17;29(6):1069-80
references: 20150896