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PMID: 20694000 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Dissecting the molecular architecture of integrin adhesion sites by cryo-electron tomography.

Nature cell biology ·Vol. 12 ·No. 9 ·2010-09-00 ·Pages 909-15

Patla I, Volberg T, Elad N, Hirschfeld-Warneken V, Grashoff C, Fässler R, Spatz JP, Geiger B, Medalia O

Abstract

Focal adhesions are integrin-based multiprotein complexes, several micrometres in diameter, that mechanically link the extracellular matrix with the termini of actin bundles. The molecular diversity of focal adhesions and their role in cell migration and matrix sensing has been extensively studied, but their ultrastructural architecture is still unknown. We present the first three-dimensional structural reconstruction of focal adhesions using cryo-electron tomography. Our analyses reveal that the membrane-cytoskeleton interaction at focal adhesions is mediated through particles located at the cell membrane and attached to actin fibres. The particles have diameters of 25 +/- 5 nm, and an average interspacing of approximately 45 nm. Treatment with the Rho-kinase inhibitor Y-27632 induces a rapid decrease in particle diameter, suggesting that they are highly mechanosensitive. Our findings clarify the internal architecture of focal adhesions at molecular resolution, and provide insights into their scaffolding and mechanosensory functions.

MeSH Terms
Actin Cytoskeleton/metabolism Amides/pharmacology Animals Cell Line Cryoelectron Microscopy/methods Fibroblasts/drug effects,ultrastructure Focal Adhesions/drug effects,metabolism,ultrastructure Integrins/metabolism Microscopy, Fluorescence Models, Molecular Particle Size Paxillin/metabolism Protein Kinase Inhibitors/pharmacology Pyridines/pharmacology Rats Stress Fibers/ultrastructure Tomography/methods Vinculin/metabolism rho-Associated Kinases/antagonists & inhibitors
Chemicals
Amides Integrins Paxillin Protein Kinase Inhibitors Pxn protein, rat Pyridines Vcl protein, rat Vinculin Y 27632 rho-Associated Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Patla Israel
Department of Life Sciences and the National Institute for Biotechnology in the Negev, Ben Gurion University of the Negev, Be'er-Sheva 84120, Israel.
Volberg Tova
Elad Nadav
Hirschfeld-Warneken Vera
Grashoff Carsten
Fässler Reinhard
Spatz Joachim P
Geiger Benjamin
Medalia Ohad
References (44)
44 references, click to expand
  1. Focal adhesions: structure and dynamics.
    Biol Cell. 2000 Oct;92(7):477-94 PMID: 11229600
  2. Organization of talin and vinculin in adhesion plaques of wet-cleaved chicken embryo fibroblasts.
    J Cell Sci. 1991 Nov;100 ( Pt 3):579-87 PMID: 1808206
  3. Focal contacts as mechanosensors: externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism.
    J Cell Biol. 2001 Jun 11;153(6):1175-86 PMID: 11402062
  4. Cryo-electron microscopy of vitrified specimens.
    Q Rev Biophys. 1988 May;21(2):129-228 PMID: 3043536
  5. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton.
    Annu Rev Cell Biol. 1988;4:487-525 PMID: 3058164
  6. A role for the juxtamembrane cytoplasm in the molecular dynamics of focal adhesions.
    PLoS One. 2009;4(1):e4304 PMID: 19172999
  7. Correlative microscopy: bridging the gap between fluorescence light microscopy and cryo-electron tomography.
    J Struct Biol. 2007 Nov;160(2):135-45 PMID: 17884579
  8. Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography.
    Science. 2002 Nov 8;298(5596):1209-13 PMID: 12424373
  9. Local call: from integrins to actin assembly.
    Trends Cell Biol. 2006 Jul;16(7):327-9 PMID: 16769214
  10. Genetic and cell biological analysis of integrin outside-in signaling.
    Genes Dev. 2009 Feb 15;23(4):397-418 PMID: 19240129
  11. Cell spreading and focal adhesion dynamics are regulated by spacing of integrin ligands.
    Biophys J. 2007 Apr 15;92(8):2964-74 PMID: 17277192
  12. Cell adhesion receptors in mechanotransduction.
    Curr Opin Cell Biol. 2008 Oct;20(5):551-6 PMID: 18583124
  13. TOM software toolbox: acquisition and analysis for electron tomography.
    J Struct Biol. 2005 Mar;149(3):227-34 PMID: 15721576
  14. Rho-stimulated contractility drives the formation of stress fibers and focal adhesions.
    J Cell Biol. 1996 Jun;133(6):1403-15 PMID: 8682874
  15. Local force and geometry sensing regulate cell functions.
    Nat Rev Mol Cell Biol. 2006 Apr;7(4):265-75 PMID: 16607289
  16. The EM Program Package: A Platform for Image Processing in Biological Electron Microscopy
    J Struct Biol. 1996 Oct;116(1):30-4 PMID: 8812976
  17. Multivariate statistical classification of noisy images (randomly oriented biological macromolecules).
    Ultramicroscopy. 1984;13(1-2):165-83 PMID: 6382731
  18. Molecular cloning of human paxillin, a focal adhesion protein phosphorylated by P210BCR/ABL.
    J Biol Chem. 1995 Mar 10;270(10):5039-47 PMID: 7534286
  19. Feedback interactions between cell-cell adherens junctions and cytoskeletal dynamics in newt lung epithelial cells.
    Mol Biol Cell. 2000 Jul;11(7):2471-83 PMID: 10888682
  20. Integrins and the actin cytoskeleton.
    Curr Opin Cell Biol. 2007 Feb;19(1):43-50 PMID: 17184985
  21. Rho-kinase dependent organization of stress fibers and focal adhesions in cultured fibroblasts.
    Genes Cells. 2007 May;12(5):623-38 PMID: 17535253
  22. Dynamics of cellular focal adhesions on deformable substrates: consequences for cell force microscopy.
    Biophys J. 2008 Jul;95(2):527-39 PMID: 18408038
  23. Focal adhesions: what's new inside.
    Dev Biol. 2006 Jun 15;294(2):280-91 PMID: 16650401
  24. Molecular complexity and dynamics of cell-matrix adhesions.
    J Cell Sci. 2001 Oct;114(Pt 20):3583-90 PMID: 11707510
  25. A new method of preparing gold probes for multiple-labeling cytochemistry.
    Eur J Cell Biol. 1985 Jul;38(1):87-93 PMID: 4029177
  26. Primary sequence of paxillin contains putative SH2 and SH3 domain binding motifs and multiple LIM domains: identification of a vinculin and pp125Fak-binding region.
    J Cell Sci. 1994 Jun;107 ( Pt 6):1583-91 PMID: 7525621
  27. Environmental sensing through focal adhesions.
    Nat Rev Mol Cell Biol. 2009 Jan;10(1):21-33 PMID: 19197329
  28. Flexible substrata for the detection of cellular traction forces.
    Trends Cell Biol. 2002 Feb;12(2):79-84 PMID: 11849971
  29. Studies of focal adhesion assembly.
    Biochem Soc Trans. 2008 Apr;36(Pt 2):263-6 PMID: 18363570
  30. Multiparametric analysis of focal adhesion formation by RNAi-mediated gene knockdown.
    J Cell Biol. 2009 Aug 10;186(3):423-36 PMID: 19667130
  31. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts.
    Cell. 1979 Mar;16(3):675-85 PMID: 222466
  32. Single myosin molecule mechanics: piconewton forces and nanometre steps.
    Nature. 1994 Mar 10;368(6467):113-9 PMID: 8139653
  33. Computer graphic display method for visualizing three-dimensional biological structures.
    Science. 1986 May 30;232(4754):1113-5 PMID: 3754654
  34. Inhibition of fibronectin binding to platelets by proteolytic fragments and synthetic peptides which support fibroblast adhesion.
    J Biol Chem. 1985 Apr 10;260(7):3931-6 PMID: 3980461
  35. Functional atlas of the integrin adhesome.
    Nat Cell Biol. 2007 Aug;9(8):858-67 PMID: 17671451
  36. The heel and toe of the cell's foot: a multifaceted approach for understanding the structure and dynamics of focal adhesions.
    Cell Motil Cytoskeleton. 2009 Nov;66(11):1017-29 PMID: 19598236
  37. Substrate-attached membranes of cultured cells isolation and characterization of ventral cell membranes and the associated cytoskeleton.
    J Mol Biol. 1981 Dec 5;153(2):361-79 PMID: 7040683
  38. Contact formation during fibroblast locomotion: involvement of membrane ruffles and microtubules.
    J Cell Biol. 1988 Mar;106(3):747-60 PMID: 3126193
  39. Imaging intracellular fluorescent proteins at nanometer resolution.
    Science. 2006 Sep 15;313(5793):1642-5 PMID: 16902090
  40. Use and properties of ROCK-specific inhibitor Y-27632.
    Methods Enzymol. 2000;325:273-84 PMID: 11036610
  41. Labeling of structural elements at the ventral plasma membrane of fibroblasts with the immunogold technique.
    J Histochem Cytochem. 1987 Apr;35(4):499-506 PMID: 2880891
  42. Adhesions of fibroblasts to substratum during contact inhibition observed by interference reflection microscopy.
    Exp Cell Res. 1975 Apr;92(1):57-62 PMID: 1169157
  43. Noise reduction in electron tomographic reconstructions using nonlinear anisotropic diffusion.
    J Struct Biol. 2001 Sep;135(3):239-50 PMID: 11722164
  44. Quantitative multicolor compositional imaging resolves molecular domains in cell-matrix adhesions.
    PLoS One. 2008 Apr 02;3(4):e1901 PMID: 18382676
Article Info
Journal
Nature cell biology
Abbr.
Nat Cell Biol
ISSN
1476-4679
DOI
10.1038/ncb2095
Published
2010-09-00
Epub
2010-00-08
Pages
909-15
Language
English
Region
England
NLM ID
100890575
Subset
IM
Grants
NIGMS NIH HHS · U54 GM64346 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1038/ncb2095 View journal details
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