|
|
|
|
|
||
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 109, Issue 5 941-952, Copyright © 1996 by Company of Biologists
JOURNAL ARTICLES |
SP Palecek, CE Schmidt, DA Lauffenburger and AF Horwitz
Department of Chemical Engineering, University of Illinois at Urbana-Champaign 61801, USA.
Cell migration is a complex process that can be considered as a repeated cycle of lamellipod extension and attachment, cytoskeletal contraction, and tail detachment. While lamellipodial and cytoskeletal phenomena are currently the focus of considerable research on cell migration, under many conditions locomotion appears to be rate-limited by events at the cell rear, especially release of cell/substratum adhesions. To study the mechanism of tail detachment, we have developed a novel experimental system that permits observation of integrin dynamics on the ventral surface of migrating fibroblasts. Photoactivatable caged fluorescein is coupled to a non-adhesion-perturbing anti-avian-beta 1 integrin subunit antibody, which labels integrins on chicken fibroblasts migrating on a laminin-coated glass coverslip. Ultraviolet light is focused through a pinhole to photoactivate the caged fluorophore in a 10-micron-diameter spot at the rear of a polarized cell. The fate of integrins initially present in this spot is monitored using a cooled CCD camera to follow the movement of fluorescent intensity as a function of time over a 2 to 3 hour period. We find that a substantial fraction of the integrins is left behind on the substratum as the cell detaches and locomotes, while another fraction collects into vesicles which are transported along the cell body as the cell migrates. As aggregates rip from the cell membrane, the integrin-cytoskeletal bonds are preferentially fractured resulting in 81 +/- 15% of the integrin remaining attached to the substratum. We additionally find that adhesions sometimes disperse into integrins which can form new adhesions at other locations in the cell. Adhesions along the cell edge can release from the substrate and translocate with the cell. They either disperse in the cell membrane, rip from the cell membrane and remain attached to the substratum, or form a new aggregate. These observations indicate that the behavior of integrins at the cell rear is much more dynamic than previously appreciated, suggesting that an important locus for regulation of motility may reside in this region.
This article has been cited by other articles:
|
|
 |
|
  M. A. Digman, C. M. Brown, A. R. Horwitz, W. W. Mantulin, and E. Gratton Paxillin Dynamics Measured during Adhesion Assembly and Disassembly by Correlation Spectroscopy Biophys. J., April 1, 2008; 94(7): 2819 - 2831. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. P. Iwanicki, T. Vomastek, R. W. Tilghman, K. H. Martin, J. Banerjee, P. B. Wedegaertner, and J. T. Parsons FAK, PDZ-RhoGEF and ROCKII cooperate to regulate adhesion movement and trailing-edge retraction in fibroblasts J. Cell Sci., March 15, 2008; 121(6): 895 - 905. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Brown, B. Hebert, D. L. Kolin, J. Zareno, L. Whitmore, A. R. Horwitz, and P. W. Wiseman Probing the integrin-actin linkage using high-resolution protein velocity mapping J. Cell Sci., December 15, 2006; 119(24): 5204 - 5214. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. B. Troyanovsky, E. P. Sokolov, and S. M. Troyanovsky Endocytosis of Cadherin from Intracellular Junctions Is the Driving Force for Cadherin Adhesive Dimer Disassembly Mol. Biol. Cell, August 1, 2006; 17(8): 3484 - 3493. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Nayal, D. J. Webb, C. M. Brown, E. M. Schaefer, M. Vicente-Manzanares, and A. R. Horwitz Paxillin phosphorylation at Ser273 localizes a GIT1-PIX-PAK complex and regulates adhesion and protrusion dynamics J. Cell Biol., May 22, 2006; 173(4): 587 - 589. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Zlatanov, T. Groth, A. Lendlein, and G. Altankov Dynamics of {beta}1-Integrins in Living Fibroblasts--Effect of Substratum Wettability Biophys. J., November 1, 2005; 89(5): 3555 - 3562. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. H. Lin, Z.-Y. Park, D. Lin, A. A. Brahmbhatt, M.-C. Rio, J. R. Yates III, and R. L. Klemke Regulation of cell migration and survival by focal adhesion targeting of Lasp-1 J. Cell Biol., May 10, 2004; 165(3): 421 - 432. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. S. K. Uchida and S. Yumura Dynamics of novel feet of Dictyostelium cells during migration J. Cell Sci., March 15, 2004; 117(8): 1443 - 1455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Griffin, A. J. Engler, T. A. Barber, K. E. Healy, H. L. Sweeney, and D. E. Discher Patterning, Prestress, and Peeling Dynamics of Myocytes Biophys. J., February 1, 2004; 86(2): 1209 - 1222. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Koka, C. L. Neudauer, X. Li, R. E. Lewis, J. B. McCarthy, and J. J. Westendorf The formin-homology-domain-containing protein FHOD1 enhances cell migration J. Cell Sci., May 1, 2003; 116(9): 1745 - 1755. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Lokuta, P. A. Nuzzi, and A. Huttenlocher Calpain regulates neutrophil chemotaxis PNAS, April 1, 2003; 100(7): 4006 - 4011. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Z. Rappoport and S. M. Simon Real-time analysis of clathrin-mediated endocytosis during cell migration J. Cell Sci., March 1, 2003; 116(5): 847 - 855. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. U. Jawhari, A. Buda, M. Jenkins, K. Shehzad, C. Sarraf, M. Noda, M. J. G. Farthing, M. Pignatelli, and J. C. Adams Fascin, an Actin-Bundling Protein, Modulates Colonic Epithelial Cell Invasiveness and Differentiation in Vitro Am. J. Pathol., January 1, 2003; 162(1): 69 - 80. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Huen, J. K. Park, L. M. Godsel, X. Chen, L. J. Bannon, E. V. Amargo, T. Y. Hudson, A. K. Mongiu, I. M. Leigh, D. P. Kelsell, et al. Intermediate filament-membrane attachments function synergistically with actin-dependent contacts to regulate intercellular adhesive strength J. Cell Biol., December 23, 2002; 159(6): 1005 - 1017. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Gao, S. Negash, H. T. Guo, D. Ledee, H.-S. Wang, and P. Zelenka CDK5 Regulates Cell Adhesion and Migration in Corneal Epithelial Cells Mol. Cancer Res., November 1, 2002; 1(1): 12 - 24. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Li, P. Butler, Y. Wang, Y. Hu, D. C. Han, S. Usami, J.-L. Guan, and S. Chien The role of the dynamics of focal adhesion kinase in the mechanotaxis of endothelial cells PNAS, March 19, 2002; 99(6): 3546 - 3551. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. G. Stupack and D. A. Cheresh ECM Remodeling Regulates Angiogenesis: Endothelial Integrins Look for New Ligands Sci. Signal., February 12, 2002; 2002(119): pe7 - pe7. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R.-i. Manabe, M. Kovalenko, D. J. Webb, and A. R. Horwitz GIT1 functions in a motile, multi-molecular signaling complex that regulates protrusive activity and cell migration J. Cell Sci., January 4, 2002; 115(7): 1497 - 1510. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. M. Munro and G. M. Odell Polarized basolateral cell motility underlies invagination and convergent extension of the ascidian notochord Development, January 1, 2002; 129(1): 13 - 24. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Laukaitis, D. J. Webb, K. Donais, and A. F. Horwitz Differential Dynamics of {alpha}5 Integrin, Paxillin, and {alpha}-Actinin during Formation and Disassembly of Adhesions in Migrating Cells J. Cell Biol., June 25, 2001; 153(7): 1427 - 1440. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kawakami, H. Tatsumi, and M. Sokabe Dynamics of integrin clustering at focal contacts of endothelial cells studied by multimode imaging microscopy J. Cell Sci., January 9, 2001; 114(17): 3125 - 3135. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Mulrooney, T. Hong, and L. B. Grabel Serine 785 phosphorylation of the {beta}1 cytoplasmic domain modulates {beta}1A-integrin-dependent functions J. Cell Sci., January 7, 2001; 114(13): 2525 - 2533. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. K. Damer and T. J. O'Halloran Spatially Regulated Recruitment of Clathrin to the Plasma Membrane during Capping and Cell Translocation Mol. Biol. Cell, June 1, 2000; 11(6): 2151 - 2159. [Abstract] [Full Text]
|
|
|
|
 |
|
 L. M. Pierini, M. A. Lawson, R. J. Eddy, B. Hendey, and F. R. Maxfield Oriented endocytic recycling of alpha 5beta 1 in motile neutrophils Blood, April 15, 2000; 95(8): 2471 - 2480. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Glading, P. Chang, D. A. Lauffenburger, and A. Wells Epidermal Growth Factor Receptor Activation of Calpain Is Required for Fibroblast Motility and Occurs via an ERK/MAP Kinase Signaling Pathway J. Biol. Chem., January 28, 2000; 275(4): 2390 - 2398. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Nishizaka, Q. Shi, and M. P. Sheetz Position-dependent linkages of fibronectin- integrin-cytoskeleton PNAS, January 18, 2000; 97(2): 692 - 697. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A Scherberich, M Campos-Toimil, P Ronde, K Takeda, and A Beretz Migration of human vascular smooth muscle cells involves serum-dependent repeated cytosolic calcium transients J. Cell Sci., January 2, 2000; 113(4): 653 - 662. [Abstract] [PDF]
|
|
|
|
 |
|
 L. B. Smilenov, A. Mikhailov, R. J. Pelham Jr., E. E. Marcantonio, and G. G. Gundersen Focal Adhesion Motility Revealed in Stationary Fibroblasts Science, November 5, 1999; 286(5442): 1172 - 1174. [Abstract] [Full Text]
|
|
|
|
 |
|
 C. Phan, A. W. McMahon, R. C. Nelson, J. F. Elliott, and A. G. Murray Activated Lymphocytes Promote Endothelial Cell Detachment from Matrix: A Role for Modulation of Endothelial Cell {beta}1 Integrin Affinity J. Immunol., October 15, 1999; 163(8): 4557 - 4563. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Ra, C Picart, H Feng, H. Sweeney, and D. Discher Muscle cell peeling from micropatterned collagen: direct probing of focal and molecular properties of matrix adhesion J. Cell Sci., January 5, 1999; 112(10): 1425 - 1436. [Abstract] [PDF]
|
|
|
|
|
|
Y. Volkov, A. Long, and D. Kelleher Inside the Crawling T Cell: Leukocyte Function-Associated Antigen-1 Cross-Linking Is Associated with Microtubule-Directed Translocation of Protein Kinase C Isoenzymes {beta}(I) and {delta} J. Immunol., December 15, 1998; 161(12): 6487 - 6495. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Cao, K. Huang, and A. F. Horwitz Identification of a Domain on the Integrin alpha 5 Subunit Implicated in Cell Spreading and Signaling J. Biol. Chem., November 27, 1998; 273(48): 31670 - 31679. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. J. PETTIT Cytosolic Free Calcium and the Cytoskeleton in the Control of Leukocyte Chemotaxis Physiol Rev, October 1, 1998; 78(4): 949 - 967. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Kamiguchi, K. E. Long, M. Pendergast, A. W. Schaefer, I. Rapoport, T. Kirchhausen, and V. Lemmon The Neural Cell Adhesion Molecule L1 Interacts with the AP-2 Adaptor and Is Endocytosed via the Clathrin-Mediated Pathway J. Neurosci., July 15, 1998; 18(14): 5311 - 5321. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Palecek, A Huttenlocher, A. Horwitz, and D. Lauffenburger Physical and biochemical regulation of integrin release during rear detachment of migrating cells J. Cell Sci., January 4, 1998; 111(7): 929 - 940. [Abstract] [PDF]
|
|
|
|
|
|
H Xie, M. Pallero, K Gupta, P Chang, M. Ware, W Witke, D. Kwiatkowski, D. Lauffenburger, J. Murphy-Ullrich, and A Wells EGF receptor regulation of cell motility: EGF induces disassembly of focal adhesions independently of the motility-associated PLCgamma signaling pathway J. Cell Sci., January 3, 1998; 111(5): 615 - 624. [Abstract] [PDF]
|
|
|
|
|
|
I. Rabinovitz and A. M. Mercurio The Integrin alpha 6beta 4 Functions in Carcinoma Cell Migration on Laminin-1 by Mediating the Formation and Stabilization of Actin-containing Motility Structures J. Cell Biol., December 29, 1997; 139(7): 1873 - 1884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Huttenlocher, S. P. Palecek, Q. Lu, W. Zhang, R. L. Mellgren, D. A. Lauffenburger, M. H. Ginsberg, and A. F. Horwitz Regulation of Cell Migration by the Calcium-dependent Protease Calpain J. Biol. Chem., December 26, 1997; 272(52): 32719 - 32722. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Li, L. H. Chow, and J. G. Pickering Cell Surface-bound Collagenase-1 and Focal Substrate Degradation Stimulate the Rear Release of Motile Vascular Smooth Muscle Cells J. Biol. Chem., November 3, 2000; 275(45): 35384 - 35392. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Han, S. Liu, D. M. Rose, D. D. Schlaepfer, H. McDonald, and M. H. Ginsberg Phosphorylation of the Integrin alpha 4 Cytoplasmic Domain Regulates Paxillin Binding J. Biol. Chem., October 26, 2001; 276(44): 40903 - 40909. [Abstract] [Full Text] [PDF]
|
|
