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Cited by 279Cell Adhesion Molecules ResearchCellular Mechanics and InteractionsProtein Kinase Regulation and GTPase Signaling

Kindlin-2 cooperates with talin to activate integrins and induces cell spreading by directly binding paxillin

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Theodosiou, Marina, Widmaier, Moritz, Böttcher, Ralph T, Rognoni, Emanuel, Veelders, Maik, Bharadwaj, Mitasha, Lambacher, Armin, Austen, Katharina +3 more·2016·eLife
DOI 10.7554/elife.10130PMC PMC4749545
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Abstract

Integrins require an activation step prior to ligand binding and signaling. How talin and kindlin contribute to these events in non-hematopoietic cells is poorly understood. Here we report that fibroblasts lacking either talin or kindlin failed to activate β1 integrins, adhere to fibronectin (FN) or maintain their integrins in a high affinity conformation induced by Mn2+. Despite compromised integrin activation and adhesion, Mn2+ enabled talin- but not kindlin-deficient cells to initiate spreading on FN. This isotropic spreading was induced by the ability of kindlin to directly bind paxillin, which in turn bound focal adhesion kinase (FAK) resulting in FAK activation and the formation of lamellipodia. Our findings show that talin and kindlin cooperatively activate integrins leading to FN binding and adhesion, and that kindlin subsequently assembles an essential signaling node at newly formed adhesion sites in a talin-independent manner.

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Research works printed but not linked: 69

  1. Askari JA Tynan CJ Webb SED Martin-Fernandez ML Ballestrem C Humphries MJ 2010 Focal adhesions are sites of integrin extension The Journal of Cell Biology 188 891 903 10.1083/jcb.200907174 20231384 PMC2845069 · DOI
  2. Azimifar SB Bottcher RT Zanivan S Grashoff C Kruger M Legate KR Mann M Fassler R 2012 Induction of membrane circular dorsal ruffles requires co-signalling of integrin-ILK-complex and EGF receptor Journal of Cell Science 125 435 448 10.1242/jcs.091652 22357970 · DOI
  3. Bachir AI Zareno J Moissoglu K Plow EF Gratton E Horwitz AR 2014 Integrin-associated complexes form hierarchically with variable stoichiometry in nascent adhesions Current Biology 24 1845 1853 10.1016/j.cub.2014.07.011 25088556 PMC4138543 · DOI
  4. Bazzoni G Shih D-T Buck CA Hemler ME 1995 Monoclonal antibody 9EG7 defines a novel 1 integrin epitope induced by soluble ligand and manganese, but inhibited by calcium Journal of Biological Chemistry 270 25570 25577 10.1074/jbc.270.43.25570 7592728 · DOI
  5. Bennett JS 2005 Structure and function of the platelet integrin IIb 3 Journal of Clinical Investigation 115 3363 3369 10.1172/JCI26989 16322781 PMC1297263 · DOI
  6. Böttcher RT Stremmel C Meves A Meyer H Widmaier M Tseng H-Y Fässler R 2012 Sorting nexin 17 prevents lysosomal degradation of β1 integrins by binding to the β1-integrin tail Nature Cell Biology 14 584 592 10.1038/ncb2501 22561348 · DOI · Paper page
  7. Bouchard V Demers M-J Thibodeau S Laquerre V Fujita N Tsuruo T Beaulieu J-F Gauthier R Vézina A Villeneuve L Vachon PH 2007 Fak/Src signaling in human intestinal epithelial cell survival and anoikis: differentiation state-specific uncoupling with the PI3-K/Akt-1 and MEK/Erk pathways Journal of Cellular Physiology 212 717 728 10.1002/jcp.21096 17443665 · DOI
  8. Brami-Cherrier K Gervasi N Arsenieva D Walkiewicz K Boutterin M-C Ortega A Leonard PG Seantier B Gasmi L Bouceba T Kadare G Girault J-A Arold ST 2014 FAK dimerization controls its kinase-dependent functions at focal adhesions The EMBO Journal 33 356 370 10.1002/embj.201386399 24480479 PMC3989642 · DOI
  9. Brown MC Perrotta JA Turner CE 1996 Identification of LIM3 as the principal determinant of paxillin focal adhesion localization and characterization of a novel motif on paxillin directing vinculin and focal adhesion kinase binding The Journal of Cell Biology 135 1109 1123 10.1083/jcb.135.4.1109 8922390 PMC2133378 · DOI
  10. Byun J Verardo MR Sumengen B Lewis GP Manjunath BS Fisher SK 2006 Automated tool for the detection of cell nuclei in digital microscopic images: application to retinal images Molecular Vision 12 949 960 16943767
  11. Calderwood DA Campbell ID Critchley DR 2013 Talins and kindlins: partners in integrin-mediated adhesion Nature Reviews Molecular Cell Biology 14 503 517 10.1038/nrm3624 23860236 PMC4116690 · DOI
  12. Chen W Lou J Zhu C 2010 Forcing switch from short- to intermediate- and long-lived states of the a domain generates LFA-1/ICAM-1 catch bonds Journal of Biological Chemistry 285 35967 35978 10.1074/jbc.M110.155770 20819952 PMC2975219 · DOI
  13. Choi CK Zareno J Digman MA Gratton E Horwitz AR 2011 Cross-correlated fluctuation analysis reveals phosphorylation-regulated paxillin-FAK complexes in nascent adhesions Biophysical Journal 100 583 592 10.1016/j.bpj.2010.12.3719 21281572 PMC3030238 · DOI
  14. Cluzel C Saltel F Lussi J Paulhe F Imhof BA Wehrle-Haller B 2005 The mechanisms and dynamics of v 3 integrin clustering in living cells The Journal of Cell Biology 171 383 392 10.1083/jcb.200503017 16247034 PMC2171205 · DOI
  15. Conti FJ Monkley SJ Wood MR Critchley DR Muller U 2009 Talin 1 and 2 are required for myoblast fusion, sarcomere assembly and the maintenance of myotendinous junctions Development 136 3597 3606 10.1242/dev.035857 19793892 PMC2761109 · DOI
  16. Coussen F Choquet D Sheetz MP Erickson HP 2002 Trimers of the fibronectin cell adhesion domain localize to actin filament bundles and undergo rearward translocation Journal of Cell Science 115 2581 2590 12045228
  17. Deakin NO Pignatelli J Turner CE 2012 Diverse roles for the paxillin family of proteins in cancer Genes & Cancer 3 362 370 10.1177/1947601912458582 23226574 PMC3513785 · DOI
  18. Deramaudt TB Dujardin D Noulet F Martin S Vauchelles R Takeda K Rondé P Parsons M 2014 Altering FAK-paxillin interactions reduces adhesion, migration and invasion processes PLoS ONE 9 e10130 10.1371/journal.pone.0092059 PMC3958421 24642576 · DOI
  19. Du X Plow EF Frelinger AL O'Toole TE Loftus JC Ginsberg MH 1991 Ligands 'activate' integrin αIIbβ3 (platelet GPIIb-IIIa) Cell 65 409 416 10.1016/0092-8674(91)90458-B 2018974 · DOI
  20. Evans R Patzak I Svensson L De Filippo K Jones K McDowall A Hogg N 2009 Integrins in immunity Journal of Cell Science 122 215 225 10.1242/jcs.019117 19118214 · DOI
  21. Fassler R Meyer M 1995 Consequences of lack of beta 1 integrin gene expression in mice Genes & Development 9 1896 1908 10.1101/gad.9.15.1896 7544313 · DOI
  22. Friedland JC Lee MH Boettiger D 2009 Mechanically activated integrin switch controls 5 1 function Science 323 642 644 10.1126/science.1168441 19179533 · DOI
  23. Friedrichs J Helenius J Muller DJ 2010 Quantifying cellular adhesion to extracellular matrix components by single-cell force spectroscopy Nature Protocols 5 1353 1361 10.1038/nprot.2010.89 20595963 · DOI
  24. Fukuda K Bledzka K Yang J Perera HD Plow EF Qin J 2014 Molecular basis of kindlin-2 binding to integrin-linked kinase pseudokinase for regulating cell adhesion Journal of Biological Chemistry 289 28363 28375 10.1074/jbc.M114.596692 25160619 PMC4192489 · DOI
  25. Gailit J Ruoslahti E 1988 Regulation of the fibronectin receptor affinity by divalent cations The Journal of Biological Chemistry 263 12927 12932 2458338
  26. Hagel M George EL Kim A Tamimi R Opitz SL Turner CE Imamoto A Thomas SM 2002 The adaptor protein paxillin is essential for normal development in the mouse and is a critical transducer of fibronectin signaling Molecular and Cellular Biology 22 901 915 10.1128/MCB.22.3.901-915.2002 11784865 PMC133539 · DOI
  27. Han J Lim CJ Watanabe N Soriani A Ratnikov B Calderwood DA Puzon-McLaughlin W Lafuente EM Boussiotis VA Shattil SJ Ginsberg MH 2006 Reconstructing and deconstructing agonist-induced activation of integrin αIIbβ3 Current Biology 16 1796 1806 10.1016/j.cub.2006.08.035 16979556 · DOI
  28. Harburger DS Bouaouina M Calderwood DA 2009 Kindlin-1 and -2 directly bind the c-terminal region of integrin cytoplasmic tails and exert integrin-specific activation effects Journal of Biological Chemistry 284 11485 11497 10.1074/jbc.M809233200 19240021 PMC2670154 · DOI · Paper page
  29. Huet-Calderwood C Brahme NN Kumar N Stiegler AL Raghavan S Boggon TJ Calderwood DA 2014 Differences in binding to the ILK complex determines kindlin isoform adhesion localization and integrin activation Journal of Cell Science 127 4308 4321 10.1242/jcs.155879 25086068 PMC4179494 · DOI
  30. Hutter JL Bechhoefer J 1993 Calibration of atomic-force microscope tips Review of Scientific Instruments 64 1868 1873 10.1063/1.1143970 · DOI
  31. Hynes RO 2002 Integrins Cell 110 673 687 10.1016/S0092-8674(02)00971-6 12297042 · DOI
  32. Kong F Garcia AJ Mould AP Humphries MJ Zhu C 2009 Demonstration of catch bonds between an integrin and its ligand The Journal of Cell Biology 185 1275 1284 10.1083/jcb.200810002 19564406 PMC2712956 · DOI
  33. Kong F Li Z Parks WM Dumbauld DW García AJ Mould AP Humphries MJ Zhu C 2013 Cyclic mechanical reinforcement of integrin–ligand interactions Molecular Cell 49 1060 1068 10.1016/j.molcel.2013.01.015 23416109 PMC3615084 · DOI
  34. Koo LY Irvine DJ Mayes AM Lauffenburger DA Griffith LG 2002 Co-regulation of cell adhesion by nanoscale RGD organization and mechanical stimulus Journal of Cell Science 115 1423 1433 11896190
  35. Lawson C Lim S-T Uryu S Chen XL Calderwood DA Schlaepfer DD 2012 FAK promotes recruitment of talin to nascent adhesions to control cell motility The Journal of Cell Biology 196 223 232 10.1083/jcb.201108078 22270917 PMC3265949 · DOI
  36. Liao Z Kato H Pandey M Cantor JM Ablooglu AJ Ginsberg MH Shattil SJ 2015 Interaction of kindlin-2 with integrin 3 promotes outside-in signaling responses by the v 3 vitronectin receptor Blood 125 1995 2004 10.1182/blood-2014-09-603035 25587038 PMC4366628 · DOI
  37. Maheshwari G Brown G Lauffenburger DA Wells A Griffith LG 2000 Cell adhesion and motility depend on nanoscale RGD clustering Journal of Cell Science 113 1677 1686 10769199
  38. Margadant C Kreft M de Groot D-J Norman JC Sonnenberg A 2012 Distinct roles of talin and kindlin in regulating integrin α5β1 function and trafficking Current Biology 22 1554 1563 10.1016/j.cub.2012.06.060 22795696 · DOI
  39. Moik D Bottcher A Makhina T Grashoff C Bulus N Zent R Fassler R 2013 Mutations in the paxillin-binding site of integrin-linked kinase (iLK) destabilize the pseudokinase domain and cause embryonic lethality in mice Journal of Biological Chemistry 288 18863 18871 10.1074/jbc.M113.470476 23658024 PMC3696662 · DOI
  40. Montanez E Ussar S Schifferer M Bosl M Zent R Moser M Fassler R 2008 Kindlin-2 controls bidirectional signaling of integrins Genes & Development 22 1325 1330 10.1101/gad.469408 18483218 PMC2377186 · DOI
  41. Moser M Nieswandt B Ussar S Pozgajova M Fässler R 2008 Kindlin-3 is essential for integrin activation and platelet aggregation Nature Medicine 14 325 330 10.1038/nm1722 18278053 · DOI
  42. Moser M Legate KR Zent R Fassler R 2009 The tail of integrins, talin, and kindlins Science 324 895 899 10.1126/science.1163865 19443776 · DOI
  43. Mould AP Akiyama SK Humphries MJ 1995 Regulation of integrin 5 1-fibronectin interactions by divalent cations: evidence for distinct classes of binding sites for Mn2+, Mg2+, and Ca2+ Journal of Biological Chemistry 270 26270 26277 10.1074/jbc.270.44.26270 7592835 · DOI
  44. Mould AP Barton SJ Askari JA Craig SE Humphries MJ 2003 Role of ADMIDAS cation-binding site in ligand recognition by integrin 5 1 Journal of Biological Chemistry 278 51622 51629 10.1074/jbc.M306655200 14532288 · DOI
  45. Nieswandt B Moser M Pleines I Varga-Szabo D Monkley S Critchley D Fassler R 2007 Loss of talin1 in platelets abrogates integrin activation, platelet aggregation, and thrombus formation in vitro and in vivo Journal of Experimental Medicine 204 3113 3118 10.1084/jem.20071827 18086864 PMC2150972 · DOI
  46. Ovesny M K i ek P Borkovec J vindrych Z Hagen GM 2014 ThunderSTORM: a comprehensive ImageJ plug-in for PALM and STORM data analysis and super-resolution imaging Bioinformatics 30 2389 2390 10.1093/bioinformatics/btu202 24771516 PMC4207427 · DOI
  47. Pankov R Cukierman E Katz BZ Matsumoto K Lin DC Lin S Hahn C Yamada KM 2000 Integrin dynamics and matrix assembly: tensin-dependent translocation of alpha5beta1 integrins promotes early fibronectin fibrillogenesis The Journal of Cell Biology 148 1075 1090 10.1083/jcb.148.5.1075 10704455 PMC2174533 · DOI · Paper page
  48. Roca-Cusachs P Gauthier NC del Rio A Sheetz MP 2009 Clustering of 5 1 integrins determines adhesion strength whereas v 3 and talin enable mechanotransduction Proceedings of the National Academy of Sciences of the United States of America 106 16245 16250 10.1073/pnas.0902818106 19805288 PMC2752568 · DOI
  49. Scheffzek K Welti S 2012 Pleckstrin homology (pH) like domains – versatile modules in protein–protein interaction platforms FEBS Letters 586 2662 2673 10.1016/j.febslet.2012.06.006 22728242 · DOI
  50. Schlaepfer DD Mitra SK Ilic D 2004 Control of motile and invasive cell phenotypes by focal adhesion kinase Biochimica Et Biophysica Acta (BBA) - Molecular Cell Research 1692 77 102 10.1016/j.bbamcr.2004.04.008 15246681 · DOI
  51. Schubert R Strohmeyer N Bharadwaj M Ramanathan SP Krieg M Friedrichs J Franz CM Muller DJ 2014 Assay for characterizing the recovery of vertebrate cells for adhesion measurements by single-cell force spectroscopy FEBS Letters 588 3639 3648 10.1016/j.febslet.2014.06.012 24928443 · DOI
  52. Shattil SJ Kim C Ginsberg MH 2010 The final steps of integrin activation: the end game Nature Reviews Molecular Cell Biology 11 288 300 10.1038/nrm2871 20308986 PMC3929966 · DOI · Paper page
  53. Spandidos A Wang X Wang H Seed B 2010 PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification Nucleic Acids Research 38 D792 D799 10.1093/nar/gkp1005 19906719 PMC2808898 · DOI
  54. Springer TA Dustin ML 2012 Integrin inside-out signaling and the immunological synapse Current Opinion in Cell Biology 24 107 115 10.1016/j.ceb.2011.10.004 22129583 PMC3294052 · DOI
  55. Takahashi S Leiss M Moser M Ohashi T Kitao T Heckmann D Pfeifer A Kessler H Takagi J Erickson HP Fassler R 2007 The RGD motif in fibronectin is essential for development but dispensable for fibril assembly The Journal of Cell Biology 178 167 178 10.1083/jcb.200703021 17591922 PMC2064432 · DOI
  56. te Riet J Helenius J Strohmeyer N Cambi A Figdor CG Muller DJ 2014 Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6 Journal of Cell Science 127 1595 1606 10.1242/jcs.141077 24496453 · DOI
  57. Thwaites T Nogueira AT Campeotto I Silva AP Grieshaber SS Carabeo RA 2014 The chlamydia effector TarP mimics the mammalian leucine-aspartic acid motif of paxillin to subvert the focal adhesion kinase during invasion Journal of Biological Chemistry 289 30426 30442 10.1074/jbc.M114.604876 25193659 PMC4215226 · DOI
  58. Ussar S Wang H-V Linder S Fässler R Moser M 2006 The kindlins: subcellular localization and expression during murine development Experimental Cell Research 312 3142 3151 10.1016/j.yexcr.2006.06.030 16876785 · DOI
  59. Ussar S Moser M Widmaier M Rognoni E Harrer C Genzel-Boroviczeny O Fässler R van Heyningen V 2008 Loss of kindlin-1 causes skin atrophy and lethal neonatal intestinal epithelial dysfunction PLoS Genetics 4 e10130 10.1371/journal.pgen.1000289 PMC2585060 19057668 · DOI
  60. van Kooyk Y Figdor CG 2000 Avidity regulation of integrins: the driving force in leukocyte adhesion Current Opinion in Cell Biology 12 542 547 10.1016/S0955-0674(00)00129-0 10978887 · DOI
  61. van de Linde S Löschberger A Klein T Heidbreder M Wolter S Heilemann M Sauer M 2011 Direct stochastic optical reconstruction microscopy with standard fluorescent probes Nature Protocols 6 991 1009 10.1038/nprot.2011.336 21720313 · DOI
  62. Wade R Bohl J Vande Pol S 2002 Paxillin null embryonic stem cells are impaired in cell spreading and tyrosine phosphorylation of focal adhesion kinase Oncogene 21 96 107 10.1038/sj.onc.1205013 11791180 · DOI
  63. Wang P Ballestrem C Streuli CH 2011 The c terminus of talin links integrins to cell cycle progression The Journal of Cell Biology 195 499 513 10.1083/jcb.201104128 22042621 PMC3206343 · DOI
  64. Winograd-Katz SE Fässler R Geiger B Legate KR 2014 The integrin adhesome: from genes and proteins to human disease Nature Reviews Molecular Cell Biology 15 273 288 10.1038/nrm3769 24651544 · DOI
  65. Xia W Springer TA 2014 Metal ion and ligand binding of integrin α 5 β 1 Proceedings of the National Academy of Sciences of the United States of America 111 17863 17868 10.1073/pnas.1420645111 25475857 PMC4273411 · DOI
  66. Ye F Petrich BG Anekal P Lefort CT Kasirer-Friede A Shattil SJ Ruppert R Moser M Fässler R Ginsberg MH 2013 The mechanism of kindlin-mediated activation of integrin αIIbβ3 Current Biology 23 2288 2295 10.1016/j.cub.2013.09.050 24210614 PMC3912999 · DOI
  67. Zhang X Jiang G Cai Y Monkley SJ Critchley DR Sheetz MP 2008 Talin depletion reveals independence of initial cell spreading from integrin activation and traction Nature Cell Biology 10 1062 1068 10.1038/ncb1765 19160486 PMC2746969 · DOI
  68. Zhang X Moore SW Iskratsch T Sheetz MP 2014 N-WASP-directed actin polymerization activates cas phosphorylation and lamellipodium spreading Journal of Cell Science 127 1394 1405 10.1242/jcs.134692 24481817 PMC3970555 · DOI
  69. Zhu J Luo B-H Xiao T Zhang C Nishida N Springer TA 2008 Structure of a complete integrin ectodomain in a physiologic resting state and activation and deactivation by applied forces Molecular Cell 32 849 861 10.1016/j.molcel.2008.11.018 19111664 PMC2758073 · DOI

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Authors
Theodosiou, M., Widmaier, M., Böttcher, R. T., Rognoni, E., Veelders, M., Bharadwaj, M., Lambacher, A., Austen, K., Müller, D. J., Zent, R., & Fässler, R.
Affiliations
  • Max Planck Institute of Biochemistry (DE)
  • ETH Zurich (CH)
  • Vanderbilt University (US)
Corresponding author
Fässler, Reinhard
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2016-01-28
Volume
5
Pages
e10130-e10130
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.10130
PMC
PMC4749545
OpenAlex
W2277501221
Cited by
279
References
2
Citations by year
2026: 12 · 2025: 23 · 2024: 16 · 2023: 21 · 2022: 39 · 2021: 33 · 2020: 41 · 2019: 34 · 2018: 10 · 2017: 28 · 2016: 22
FWCI
29.55
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Queen Mary Research Online (Queen Mary University of London) (submittedVersion, cc-by)
  • Repository for Publications and Research Data (ETH Zurich) (submittedVersion, cc-by)
  • ETH Zürich Research Collection (cc-by)
Funders
  • Veterans Affair Merit Award (1I01BX002196)
  • NIH Office of the Director (RO1-DK083187, RO1-DK075594, R01-DK069221)
  • European Research Council (322652)
  • Deutsche Forschungsgemeinschaft (SFB-863)
  • Max-Planck-Gesellschaft
Topics
  • Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
Kindlin-2, talin, integrin activation, beta1 integrin, paxillin, focal adhesion kinase, cell spreading, fibronectin adhesion, lamellipodia, focal adhesions, fibroblasts, integrin signaling
MeSH terms
Animals, Cell Adhesion, Cell Line, Cell Movement, Cytoskeletal Proteins, Fibroblasts, Manganese, Muscle Proteins, Protein Binding, Talin, Integrin beta1, Mice, Focal Adhesion Protein-Tyrosine Kinases, Paxillin

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