The Journal of Cell Biology, Vol 129, 473-487, Copyright © 1995 by The Rockefeller University Press

ARTICLES

A recombinant tail-less integrin beta 4 subunit disrupts hemidesmosomes, but does not suppress alpha 6 beta 4-mediated cell adhesion to laminins

L Spinardi, S Einheber, T Cullen, TA Milner and FG Giancotti
Department of Pathology, New York University School of Medicine, New York 10016, USA.

To examine the function of the alpha 6 beta 4 integrin we have determined its ligand-binding ability and overexpressed two potentially dominant negative mutant beta 4 subunits, lacking either the cytoplasmic or extracellular domain, in bladder epithelial 804G cells. The results of cell adhesion and radioligand-binding assays showed that alpha 6 beta 4 is a receptor for several laminin isoforms, including laminin 1, 2, 4, and 5. Overexpression of the tail-less or head-less mutant beta 4 subunit did not suppress alpha 6 beta 4-mediated adhesion to laminins, as both types of transfectants adhered to these ligands in the presence of blocking anti-beta 1 antibodies as well as the controls. However, immunofluorescence experiments indicated that the endogenous alpha 6 beta 4 integrin and other hemidesmosomal markers were not concentrated in hemidesmosomes in cells overexpressing tail- less beta 4, while the distribution of these molecules was not altered in cells overexpressing the head-less subunit. Electron microscopic studies confirmed that cells overexpressing tail-less beta 4 had a drastically reduced number of hemidesmosomes, while cells expressing the head-less subunit had a normal number of these structures. Thus, expression of a tail-less, but not a head-less mutant beta 4 subunit leads to a dominant negative effect on hemidesmosome assembly without suppressing initial adhesion to laminins. We conclude that the alpha 6 beta 4 integrin binds to several laminins and plays an essential role in the assembly and/or stability of hemidesmosomes, that alpha 6 beta 4- mediated adhesion and hemidesmosome assembly have distinct requirements, and that it is possible to use a dominant negative approach to selectively interfere with a specific function of an integrin.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Friedland, J. C., Lakins, J. N., Kazanietz, M. G., Chernoff, J., Boettiger, D., Weaver, V. M. (2007). {alpha}6beta4 integrin activates Rac-dependent p21-activated kinase 1 to drive NF-{kappa}B-dependent resistance to apoptosis in 3D mammary acini. J. Cell Sci. 120: 3700-3712 [Abstract] [Full Text]  
• Takkunen, M., Grenman, R., Hukkanen, M., Korhonen, M., Garcia de Herreros, A., Virtanen, I. (2006). Snail-dependent and -independent Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Cells. J. Histochem. Cytochem. 54: 1263-1275 [Abstract] [Full Text]  
• Nikolopoulos, S. N., Blaikie, P., Yoshioka, T., Guo, W., Puri, C., Tacchetti, C., Giancotti, F. G. (2005). Targeted Deletion of the Integrin {beta}4 Signaling Domain Suppresses Laminin-5-Dependent Nuclear Entry of Mitogen-Activated Protein Kinases and NF-{kappa}B, Causing Defects in Epidermal Growth and Migration. Mol. Cell. Biol. 25: 6090-6102 [Abstract] [Full Text]  
• Diaz, L. K., Zhou, X., Welch, K., Sahin, A., Gilcrease, M. Z. (2004). Chromogenic In Situ Hybridization for {alpha}6{beta}4 Integrin in Breast Cancer: Correlation with Protein Expression. J. Mol. Diagn. 6: 10-15 [Abstract] [Full Text]  
• Lenander, C, Roblick, U J, Habermann, J K, Ost, A, Tryggvason, K, Auer, G (2003). Laminin 5 {gamma}2 chain expression: a marker of early invasiveness in colorectal adenomas. Mol. Pathol. 56: 342-346 [Abstract] [Full Text]  
• Tsuruta, D., Hopkinson, S. B., Lane, K. D., Werner, M. E., Cryns, V. L., Jones, J. C. R. (2003). Crucial Role of the Specificity-determining Loop of the Integrin {beta}4 Subunit in the Binding of Cells to Laminin-5 and Outside-in Signal Transduction. J. Biol. Chem. 278: 38707-38714 [Abstract] [Full Text]  
• Yeh, M.-G., Ziober, B. L., Liu, B., Lipkina, G., Vizirianakis, I. S., Kramer, R. H. (2003). The {beta}1 Cytoplasmic Domain Regulates the Laminin-binding Specificity of the {alpha}7X1 Integrin. Mol. Biol. Cell 14: 3507-3518 [Abstract] [Full Text]  
• Miralem, T., Avraham, H. K. (2003). Extracellular Matrix Enhances Heregulin-Dependent BRCA1 Phosphorylation and Suppresses BRCA1 Expression through Its C Terminus. Mol. Cell. Biol. 23: 579-593 [Abstract] [Full Text]  
• Morena, A., Riccioni, S., Marchetti, A., Polcini, A. T., Mercurio, A. M., Blandino, G., Sacchi, A., Falcioni, R. (2002). Expression of the beta 4 integrin subunit induces monocytic differentiation of 32D/v-Abl cells. Blood 100: 96-106 [Abstract] [Full Text]  
• Alt, A., Ohba, M., Li, L., Gartsbein, M., Belanger, A., Denning, Mitchell. F., Kuroki, T., Yuspa, S. H., Tennenbaum, T. (2001). Protein Kinase C{{delta}}-mediated Phosphorylation of {{alpha}}6{beta}4 Is Associated with Reduced Integrin Localization to the Hemidesmosome and Decreased Keratinocyte Attachment. Cancer Res. 61: 4591-4598 [Abstract] [Full Text]  
• Hintermann, E., Bilban, M., Sharabi, A., Quaranta, V. (2001). Inhibitory Role of {alpha}6{beta}4-Associated Erbb-2 and Phosphoinositide 3-Kinase in Keratinocyte Haptotactic Migration Dependent on {alpha}3{beta}1 Integrin. JCB 153: 465-478 [Abstract] [Full Text]  
• Bhol, K. C., Dans, M. J., Simmons, R. K., Foster, C. S., Giancotti, F. G., Ahmed, A. R. (2000). The Autoantibodies to {alpha}6{beta}4 Integrin of Patients Affected by Ocular Cicatricial Pemphigoid Recognize Predominantly Epitopes Within the Large Cytoplasmic Domain of Human {beta}4. J. Immunol. 165: 2824-2829 [Abstract] [Full Text]  
• Gambaletta, D., Marchetti, A., Benedetti, L., Mercurio, A. M., Sacchi, A., Falcioni, R. (2000). Cooperative Signaling between alpha 6beta 4 Integrin and ErbB-2 Receptor Is Required to Promote Phosphatidylinositol 3-Kinase-dependent Invasion. J. Biol. Chem. 275: 10604-10610 [Abstract] [Full Text]  
• Hopkinson, S. B., Jones, J. C. R. (2000). The N Terminus of the Transmembrane Protein BP180 Interacts with the N-terminal Domain of BP230, Thereby Mediating Keratin Cytoskeleton Anchorage to the Cell Surface at the Site of the Hemidesmosome. Mol. Biol. Cell 11: 277-286 [Abstract] [Full Text]  
• Basora, N., Herring-Gillam, F. E., Boudreau, F., Perreault, N., Pageot, L.-P., Simoneau, M., Bouatrouss, Y., Beaulieu, J.-F. (1999). Expression of Functionally Distinct Variants of the beta 4A Integrin Subunit in Relation to the Differentiation State in Human Intestinal Cells. J. Biol. Chem. 274: 29819-29825 [Abstract] [Full Text]  
• Muschler, J., Lochter, A., Roskelley, C. D., Yurchenco, P., Bissell, M. J. (1999). Division of Labor among the alpha 6beta 4 Integrin, beta 1 Integrins, and an E3 Laminin Receptor to Signal Morphogenesis and beta -Casein Expression in Mammary Epithelial Cells. Mol. Biol. Cell 10: 2817-2828 [Abstract] [Full Text]  
• Crowe, D.L., Milo, G.E., Shuler, C.E. (1999). Keratin 19 Downregulation by Oral Squamous Cell Carcinoma Lines Increases Invasive Potential. JDR 78: 1256-1263 [Abstract]  
• Pinkstaff, J. K., Detterich, J., Lynch, G., Gall, C. (1999). Integrin Subunit Gene Expression Is Regionally Differentiated in Adult Brain. J. Neurosci. 19: 1541-1556 [Abstract] [Full Text]  
• van der Neut, R, Cachaco, A., Thorsteinsdottir, S, Janssen, H, Prins, D, Bulthuis, J, van der Valk, M, Calafat, J, Sonnenberg, A (1999). Partial rescue of epithelial phenotype in integrin beta4 null mice by a keratin-5 promoter driven human integrin beta4 transgene. J. Cell Sci. 112: 3911-3922 [Abstract]  
• Balzar, M., Bakker, H. A. M., Briaire-de-Bruijn, I. H., Fleuren, G. J., Warnaar, S. O., Litvinov, S. V. (1998). Cytoplasmic Tail Regulates the Intercellular Adhesion Function of the Epithelial Cell Adhesion Molecule. Mol. Cell. Biol. 18: 4833-4843 [Abstract] [Full Text]  
• Schaapveld, R. Q.J., Borradori, L., Geerts, D., van Leusden, M. R., Kuikman, I., Nievers, M. G., Niessen, C. M., Steenbergen, R. D.M., Snijders, P. J.F., Sonnenberg, A. (1998). Hemidesmosome Formation Is Initiated by the {beta}4 Integrin Subunit, Requires Complex Formation of {beta}4 and HD1/Plectin, and Involves a Direct Interaction between {beta}4 and the Bullous Pemphigoid Antigen 180. JCB 142: 271-284 [Abstract] [Full Text]  
• Hirako, Y., Usukura, J., Uematsu, J., Hashimoto, T., Kitajima, Y., Owaribe, K. (1998). Cleavage of BP180, a 180-kDa Bullous Pemphigoid Antigen, Yields a 120-kDa Collagenous Extracellular Polypeptide. J. Biol. Chem. 273: 9711-9717 [Abstract] [Full Text]  
• Homan, S., Mercurio, A., LaFlamme, S. (1998). Endothelial cells assemble two distinct alpha6beta4-containing vimentin-associated structures: roles for ligand binding and the beta4 cytoplasmic tail. J. Cell Sci. 111: 2717-2728 [Abstract]  
• Nievers, M., Schaapveld, R., Oomen, L., Fontao, L, Geerts, D, Sonnenberg, A (1998). Ligand-independent role of the beta 4 integrin subunit in the formation of hemidesmosomes. J. Cell Sci. 111: 1659-1672 [Abstract]  
• Biffo, S., Sanvito, F., Costa, S., Preve, L., Pignatelli, R., Spinardi, L., Marchisio, P. C. (1997). Isolation of a Novel beta 4 Integrin-binding Protein (p27BBP) Highly Expressed in Epithelial Cells. J. Biol. Chem. 272: 30314-30321 [Abstract] [Full Text]  
• Colognato, H., MacCarrick, M., O'Rear, J. J., Yurchenco, P. D. (1997). The Laminin alpha 2-Chain Short Arm Mediates Cell Adhesion through Both the alpha 1beta 1 and alpha 2beta 1 Integrins. J. Biol. Chem. 272: 29330-29336 [Abstract] [Full Text]  
• Wagner, S., Tagaya, M., Koziol, J. A., Quaranta, V., del Zoppo, G. J. (1997). Rapid Disruption of an Astrocyte Interaction With the Extracellular Matrix Mediated by Integrin {alpha}6ß4 During Focal Cerebral Ischemia/Reperfusion. Stroke 28: 858-865 [Abstract] [Full Text]  
• Stahl, S, Weitzman, S, Jones, J. (1997). The role of laminin-5 and its receptors in mammary epithelial cell branching morphogenesis. J. Cell Sci. 110: 55-63 [Abstract]  
• Sanchez-Aparicio, P, Martinez de Velasco, A., Niessen, C., Borradori, L, Kuikman, I, Hulsman, E., Fassler, R, Owaribe, K, Sonnenberg, A (1997). The subcellular distribution of the high molecular mass protein, HD1, is determined by the cytoplasmic domain of the integrin beta 4 subunit. J. Cell Sci. 110: 169-178 [Abstract]  
• Yao, C.-C., Ziober, B. L., Squillace, R. M., Kramer, R. H. (1996). alpha 7 Integrin Mediates Cell Adhesion and Migration on Specific Laminin Isoforms. J. Biol. Chem. 271: 25598-25603 [Abstract] [Full Text]  
• Gagnoux-Palacios, L., Vailly, J., Durand-Clement, M., Wagner, E., Ortonne, J.-P., Meneguzzi, G. (1996). Functional Re-expression of Laminin-5 in Laminin-gamma 2-deficient Human Keratinocytes Modifies Cell Morphology, Motility, and Adhesion. J. Biol. Chem. 271: 18437-18444 [Abstract] [Full Text]  
• Hirako, Y., Usukura, J., Nishizawa, Y., Owaribe, K. (1996). Demonstration of the Molecular Shape of BP180, a 180-kDa Bullous Pemphigoid Antigen and Its Potential for Trimer Formation. J. Biol. Chem. 271: 13739-13745 [Abstract] [Full Text]  
• Falk, M, Salmivirta, K, Durbeej, M, Larsson, E, Ekblom, M, Vestweber, D, Ekblom, P (1996). Integrin alpha 6B beta 1 is involved in kidney tubulogenesis in vitro. J. Cell Sci. 109: 2801-2810 [Abstract]  
• Yao, C., Ziober, B., Sutherland, A., Mendrick, D., Kramer, R. (1996). Laminins promote the locomotion of skeletal myoblasts via the alpha 7 integrin receptor. J. Cell Sci. 109: 3139-3150 [Abstract]  
• Baker, S., Hopkinson, S., Fitchmun, M, Andreason, G., Frasier, F, Plopper, G, Quaranta, V, Jones, J. (1996). Laminin-5 and hemidesmosomes: role of the alpha 3 chain subunit in hemidesmosome stability and assembly. J. Cell Sci. 109: 2509-2520 [Abstract]  
• Niessen, C., van der Raaij-Helmer, M., Hulsman, E., van der Neut, R, Jonkman, M., Sonnenberg, A (1996). Deficiency of the integrin beta 4 subunit in junctional epidermolysis bullosa with pyloric atresia: consequences for hemidesmosome formation and adhesion properties. J. Cell Sci. 109: 1695-1706 [Abstract]  
• Giancotti, F. (1996). Signal transduction by the alpha 6 beta 4 integrin: charting the path between laminin binding and nuclear events. J. Cell Sci. 109: 1165-1172  
• Clarke, A. S., Lotz, M. M., Chao, C., Mercurio, A. M. (1995). Activation of the p21 Pathway of Growth Arrest and Apoptosis by the beta(4) Integrin Cytoplasmic Domain. J. Biol. Chem. 270: 22673-22676 [Abstract] [Full Text]  
• Barberis, L., Wary, K. K., Fiucci, G., Liu, F., Hirsch, E., Brancaccio, M., Altruda, F., Tarone, G., Giancotti, F. G. (2000). Distinct Roles of the Adaptor Protein Shc and Focal Adhesion Kinase in Integrin Signaling to ERK. J. Biol. Chem. 275: 36532-36540 [Abstract] [Full Text]  
• Abdel-Ghany, M., Cheng, H.-C., Elble, R. C., Pauli, B. U. (2001). The Breast Cancer beta 4 Integrin and Endothelial Human CLCA2 Mediate Lung Metastasis. J. Biol. Chem. 276: 25438-25446 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents