The Journal of Cell Biology, Vol 110, 175-184, Copyright © 1990 by The Rockefeller University Press

ARTICLES

Expression and function of chicken integrin beta 1 subunit and its cytoplasmic domain mutants in mouse NIH 3T3 cells

Y Hayashi, B Haimovich, A Reszka, D Boettiger and A Horwitz
Department of Cell and Structural Biology, University of Illinois, Urbana 61801.

Chicken integrin beta 1 cDNA and its site-directed mutants were cloned into a mammalian expression vector and introduced into mouse NIH 3T3 cells. Stable transfectants expressing the chicken beta 1 subunit or its site-directed mutants were identified by immunostaining with antibodies specific for the chicken integrin beta 1 subunit. The chicken beta 1 proteins were expressed predominately in the endoplasmic reticulum of transfectants and to a lesser degree in the plasma membrane. Immunoblots and immunoprecipitations, using anti-chicken integrin antibodies, revealed three different sizes of the chicken subunit (90, 95, and 120 kD) and a mouse 140-kD alpha subunit. Immunoprecipitations of the cell surface receptors showed only two peptides, an 120-kD beta 1 and an 140-kD alpha subunit. Antibodies perturbing mouse and chicken integrin-specific cell adhesions were used to demonstrate that the chimeric receptors functioned in adhesion to both laminin and fibronectin. Immunofluorescent staining with antibodies specific for either the chicken or mouse receptors showed that both the wild type and the chimeric receptors localized in focal contacts. Several mutations in the cytoplasmic domain were synthesized and used in the transfection experiments. In one mutant the tyrosine (Tyr 788) in the consensus sequence for phosphorylation was replaced by a phenylalanine. In another the lysine (Lys 757) at the end of the membrane spanning region was replaced by a leucine. Both of these mutants formed dimers with mouse alpha subunits, participated in adhesion, localized in focal contacts, and displayed biological properties indistinguishable from the wild-type transfection. In contrast, mutants containing deletions greater than 5-15 amino acids nearest the carboxyl end in the cytoplasmic domain neither promoted adhesion nor localized in focal contacts. They did, however, form heterodimers that were expressed on the cell surface.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• McKillop, W. M., Barrett, J. W., Pasternak, S. H., Chan, B. M. C., Dekaban, G. A. (2009). The extracellular domain of CD11d regulates its cell surface expression. J. Leukoc. Biol. 86: 851-862 [Abstract] [Full Text]  
• Green, J. A., Berrier, A. L., Pankov, R., Yamada, K. M. (2009). {beta}1 Integrin Cytoplasmic Domain Residues Selectively Modulate Fibronectin Matrix Assembly and Cell Spreading through Talin and Akt-1. J. Biol. Chem. 284: 8148-8159 [Abstract] [Full Text]  
• Wiedemann, A., Patel, J. C., Lim, J., Tsun, A., van Kooyk, Y., Caron, E. (2006). Two distinct cytoplasmic regions of the {beta}2 integrin chain regulate RhoA function during phagocytosis.. JCB 172: 1069-1079 [Abstract] [Full Text]  
• Armulik, A., Velling, T., Johansson, S. (2004). The Integrin {beta}1 Subunit Transmembrane Domain Regulates Phosphatidylinositol 3-Kinase-dependent Tyrosine Phosphorylation of Crk-associated Substrate. Mol. Biol. Cell 15: 2558-2567 [Abstract] [Full Text]  
• Kaariainen, M., Nissinen, L., Kaufman, S., Sonnenberg, A., Jarvinen, M., Heino, J., Kalimo, H. (2002). Expression of {alpha}7{beta}1 Integrin Splicing Variants during Skeletal Muscle Regeneration. Am. J. Pathol. 161: 1023-1031 [Abstract] [Full Text]  
• Miao, H., Li, S., Hu, Y.-L., Yuan, S., Zhao, Y., Chen, B. P. C., Puzon-McLaughlin, W., Tarui, T., Shyy, J. Y.-J., Takada, Y., Usami, S., Chien, S. (2002). Differential regulation of Rho GTPases by {beta}1 and {beta}3 integrins: the role of an extracellular domain of integrin in intracellular signaling. J. Cell Sci. 115: 2199-2206 [Abstract] [Full Text]  
• Leong, K. G., Hu, X., Li, L., Noseda, M., Larrivee, B., Hull, C., Hood, L., Wong, F., Karsan, A. (2002). Activated Notch4 Inhibits Angiogenesis: Role of {beta}1-Integrin Activation. Mol. Cell. Biol. 22: 2830-2841 [Abstract] [Full Text]  
• Chang, D. D., Hoang, B. Q., Liu, J., Springer, T. A. (2002). Molecular Basis for Interaction between Icap1alpha PTB Domain and beta 1 Integrin. J. Biol. Chem. 277: 8140-8145 [Abstract] [Full Text]  
• Gustavsson, A., Armulik, A., Brakebusch, C., Fassler, R., Johansson, S., Fallman, M. (2002). Role of the {beta}1-integrin cytoplasmic tail in mediating invasin-promoted internalization of Yersinia. J. Cell Sci. 115: 2669-2678 [Abstract] [Full Text]  
• Maitra, N., Flink, I. L., Bahl, J. J., Morkin, E. (2000). Expression of {alpha} and {beta} integrins during terminal differentiation of cardiomyocytes. Cardiovasc Res 47: 715-725 [Abstract] [Full Text]  
• Levy, L., Broad, S., Diekmann, D., Evans, R. D., Watt, F. M. (2000). beta 1 Integrins Regulate Keratinocyte Adhesion and Differentiation by Distinct Mechanisms. Mol. Biol. Cell 11: 453-466 [Abstract] [Full Text]  
• Liu, S, Calderwood, D., Ginsberg, M. (2000). Integrin cytoplasmic domain-binding proteins. J. Cell Sci. 113: 3563-3571 [Abstract]  
• Armulik, A., Nilsson, I., von Heijne, G., Johansson, S. (1999). Determination of the Border between the Transmembrane and Cytoplasmic Domains of Human Integrin Subunits. J. Biol. Chem. 274: 37030-37034 [Abstract] [Full Text]  
• van Kooyk, Y., van Vliet, S. J., Figdor, C. G. (1999). The Actin Cytoskeleton Regulates LFA-1 Ligand Binding through Avidity Rather than Affinity Changes. J. Biol. Chem. 274: 26869-26877 [Abstract] [Full Text]  
• Meredith, J. E. Jr., Kiosses, W. B., Takada, Y., Schwartz, M. A. (1999). Mutational Analysis of Cell Cycle Inhibition by Integrin beta 1C. J. Biol. Chem. 274: 8111-8116 [Abstract] [Full Text]  
• Zhang, X. A., Hemler, M. E. (1999). Interaction of the Integrin beta 1 Cytoplasmic Domain with ICAP-1 Protein. J. Biol. Chem. 274: 11-19 [Abstract] [Full Text]  
• Reddy, K. B., Gascard, P., Price, M. G., Negrescu, E. V., Fox, J. E. B. (1998). Identification of an Interaction between the M-band Protein Skelemin and beta -Integrin Subunits. COLOCALIZATION OF A SKELEMIN-LIKE PROTEIN WITH beta 1- and beta 3-INTEGRINS IN NON-MUSCLE CELLS. J. Biol. Chem. 273: 35039-35047 [Abstract] [Full Text]  
• Loo, D. T., Kanner, S. B., Aruffo, A. (1998). Filamin Binds to the Cytoplasmic Domain of the beta 1-Integrin. IDENTIFICATION OF AMINO ACIDS RESPONSIBLE FOR THIS INTERACTION. J. Biol. Chem. 273: 23304-23312 [Abstract] [Full Text]  
• Nagel, W., Zeitlmann, L., Schilcher, P., Geiger, C., Kolanus, J., Kolanus, W. (1998). Phosphoinositide 3-OH Kinase Activates the beta 2 Integrin Adhesion Pathway and Induces Membrane Recruitment of Cytohesin-1. J. Biol. Chem. 273: 14853-14861 [Abstract] [Full Text]  
• Martin-Bermudo, M. D., Dunin-Borkowski, O. M., Brown, N. H. (1998). Modulation of Integrin Activity is Vital for Morphogenesis. JCB 141: 1073-1081 [Abstract] [Full Text]  
• Schaffner-Reckinger, E., Gouon, V., Melchior, C., Plancon, S., Kieffer, N. (1998). Distinct Involvement of beta 3 Integrin Cytoplasmic Domain Tyrosine Residues 747 and 759 in Integrin-mediated Cytoskeletal Assembly and Phosphotyrosine Signaling. J. Biol. Chem. 273: 12623-12632 [Abstract] [Full Text]  
• Huttenlocher, A., Lakonishok, M., Kinder, M., Wu, S., Truong, T., Knudsen, K. A., Horwitz, A. F. (1998). Integrin and Cadherin Synergy Regulates Contact Inhibition of Migration and Motile Activity. JCB 141: 515-526 [Abstract] [Full Text]  
• Sakai, T., Zhang, Q., Fassler, R., Mosher, D. F. (1998). Modulation of {beta}1A Integrin Functions by Tyrosine Residues in the {beta}1 Cytoplasmic Domain. JCB 141: 527-538 [Abstract] [Full Text]  
• Wennerberg, K, Fassler, R, Warmegard, B, Johansson, S (1998). Mutational analysis of the potential phosphorylation sites in the cytoplasmic domain of integrin beta1A. Requirement for threonines 788-789 in receptor activation. J. Cell Sci. 111: 1117-1126 [Abstract]  
• Li, X, Graner, M., Williams, E., Roote, C., Bunch, T., Zusman, S (1998). Requirements for the cytoplasmic domain of the alphaPS1, alphaPS2 and betaPS integrin subunits during Drosophila development. Development 125: 701-711 [Abstract]  
• Agius, E., Cochard, P. (1998). Comparison of Neurite Outgrowth Induced by Intact and Injured Sciatic Nerves: A Confocal and Functional Analysis. J. Neurosci. 18: 328-338 [Abstract] [Full Text]  
• Belkin, A. M., Retta, S. F., Pletjushkina, O. Y., Balzac, F., Silengo, L., Fassler, R., Koteliansky, V. E., Burridge, K., Tarone, G. (1997). Muscle {beta}1D Integrin Reinforces the Cytoskeleton-Matrix Link: Modulation of Integrin Adhesive Function by Alternative Splicing. JCB 139: 1583-1595 [Abstract] [Full Text]  
• Sagawa, K., Kimura, T., Swieter, M., Siraganian, R. P. (1997). The Protein-tyrosine Phosphatase SHP-2 Associates with Tyrosine-phosphorylated Adhesion Molecule PECAM-1 (CD31). J. Biol. Chem. 272: 31086-31091 [Abstract] [Full Text]  
• Simon, K. O., Nutt, E. M., Abraham, D. G., Rodan, G. A., Duong, L. T. (1997). The alpha vbeta 3 Integrin Regulates alpha 5beta 1-mediated Cell Migration toward Fibronectin. J. Biol. Chem. 272: 29380-29389 [Abstract] [Full Text]  
• Lyman, S., Gilmore, A., Burridge, K., Gidwitz, S., White II, G. C. (1997). Integrin-mediated Activation of Focal Adhesion Kinase Is Independent of Focal Adhesion Formation or Integrin Activation. STUDIES WITH ACTIVATED AND INHIBITORY beta 3 CYTOPLASMIC DOMAIN MUTANTS. J. Biol. Chem. 272: 22538-22547 [Abstract] [Full Text]  
• Sagawa, K., Swaim, W., Zhang, J., Unsworth, E., Siraganian, R. P. (1997). Aggregation of the High Affinity IgE Receptor Results in the Tyrosine Phosphorylation of the Surface Adhesion Protein PECAM-1 (CD31). J. Biol. Chem. 272: 13412-13418 [Abstract] [Full Text]  
• Tahiliani, P. D., Singh, L., Auer, K. L., LaFlamme, S. E. (1997). The Role of Conserved Amino Acid Motifs within the Integrin beta 3 Cytoplasmic Domain in Triggering Focal Adhesion Kinase Phosphorylation. J. Biol. Chem. 272: 7892-7898 [Abstract] [Full Text]  
• Cruz, M., Dalgard, C., Ignatius, M. (1997). Functional partitioning of beta1 integrins revealed by activating and inhibitory mAbs. J. Cell Sci. 110: 2647-2659 [Abstract]  
• Vignoud, L, Albiges-Rizo, C, Frachet, P, Block, M. (1997). NPXY motifs control the recruitment of the alpha5beta1 integrin in focal adhesions independently of the association of talin with the beta1 chain. J. Cell Sci. 110: 1421-1430 [Abstract]  
• Puzon-McLaughlin, W., Yednock, T. A., Takada, Y. (1996). Regulation of Conformation and Ligand Binding Function of Integrin alpha 5beta 1 by the beta 1 Cytoplasmic Domain. J. Biol. Chem. 271: 16580-16585 [Abstract] [Full Text]  
• Cunningham, J. G., Meyer, S. C., Fox, J. E. B. (1996). The Cytoplasmic Domain of the alpha-Subunit of Glycoprotein (GP) Ib Mediates Attachment of the Entire GP Ib-IX Complex to the Cytoskeleton and Regulates von Willebrand Factor-induced Changes in Cell Morphology. J. Biol. Chem. 271: 11581-11587 [Abstract] [Full Text]  
• Van Nhieu, G. T., Krukonis, E. S., Reszka, A. A., Horwitz, A. F., Isberg, R. R. (1996). Mutations in the Cytoplasmic Domain of the Integrin beta(1) Chain Indicate a Role for Endocytosis Factors in Bacterial Internalization. J. Biol. Chem. 271: 7665-7672 [Abstract] [Full Text]  
• Bazzoni, G., Shih, D.-T., Buck, C. A., Hemler, M. E. (1995). Monoclonal Antibody 9EG7 Defines a Novel beta(1) Integrin Epitope Induced by Soluble Ligand and Manganese, but Inhibited by Calcium. J. Biol. Chem. 270: 25570-25577 [Abstract] [Full Text]  
• Hughes, P. E., O'Toole, T. E., Ylänne, J., Shattil, S. J., Ginsberg, M. H. (1995). The Conserved Membrane-proximal Region of an Integrin Cytoplasmic Domain Specifies Ligand Binding Affinity. J. Biol. Chem. 270: 12411-12417 [Abstract] [Full Text]  
• Ylänne, J., Huuskonen, J., O'Toole, T. E., Ginsberg, M. H., Virtanen, I., Gahmberg, C. G. (1995). Mutation of the Cytoplasmic Domain of the Integrin [IMAGE][IMAGE]Subunit. J. Biol. Chem. 270: 9550-9557 [Abstract] [Full Text]  
• Cattelino, A, Longhi, R, de Curtis, I (1995). Differential distribution of two cytoplasmic variants of the alpha 6 beta 1 integrin laminin receptor in the ventral plasma membrane of embryonic fibroblasts. J. Cell Sci. 108: 3067-3078 [Abstract]  
• McDonald, K., Lakonishok, M, Horwitz, A. (1995). Alpha v and alpha 3 integrin subunits are associated with myofibrils during myofibrillogenesis. J. Cell Sci. 108: 2573-2581 [Abstract]  
• McDonald, K., Lakonishok, M, Horwitz, A. (1995). Alpha v and alpha 3 integrin subunits are associated with myofibrils during myofibrillogenesis. J. Cell Sci. 108: 975-983 [Abstract]  
• Grinblat, Y, Zusman, S, Yee, G, Hynes, R., Kafatos, F. (1994). Functions of the cytoplasmic domain of the beta PS integrin subunit during Drosophila development. Development 120: 91-102 [Abstract]  
• Kinoshita, Y., Kinoshita, C., Heuer, J. G., Bothwell, M. (1993). Basic fibroblast growth factor promotes adhesive interactions of neuroepithelial cells from chick neural tube with extracellular matrix proteins in culture. Development 119: 943-956 [Abstract]  
• Bao, Z., Lakonishok, M, Kaufman, S, Horwitz, A. (1993). Alpha 7 beta 1 integrin is a component of the myotendinous junction on skeletal muscle. J. Cell Sci. 106: 579-589 [Abstract]  
• Tranqui, L, Usson, Y, Marie, C, Block, M. (1993). Adhesion of CHO cells to fibronectin is mediated by functionally and structurally distinct adhesion plaques. J. Cell Sci. 106: 377-387 [Abstract]  
• Adams, J., Watt, F. (1993). Regulation of development and differentiation by the extracellular matrix. Development 117: 1183-1198  
• Geiger, B., Salomon, D., Takeichi, M., Hynes, R. O. (1992). A chimeric N-cadherin/beta 1-integrin receptor which localizes to both cell-cell and cell-matrix adhesions. J. Cell Sci. 103: 943-951 [Abstract]  
• Gilmore, A. P., Jackson, P., Waites, G. T., Critchley, D. R. (1992). Further characterisation of the talin-binding site in the cytoskeletal protein vinculin. J. Cell Sci. 103: 719-731 [Abstract]  
• Yamada, K.M., Aota, S., Akiyama, S.K., LaFlamme, S.E. (1992). Mechanisms of Fibronectin and Integrin Function during Cell Adhesion and Migration. Cold Spring Harb Symp Quant Biol 57: 203-212 [Abstract]  
• Hemler, M.E., Kassner, P.D., Chan, B.M.C. (1992). Functional Roles for Integrin {alpha} Subunit Cytoplasmic Domains. Cold Spring Harb Symp Quant Biol 57: 213-220 [Abstract]  
• Geiger, B., Ayalon, O., Ginsberg, D., Volberg, T., Rodruguez Fernandez, J.L., Yarden, Y., Ben-Ze'ev, A. (1992). Cytoplasmic Control of Cell Adhesion. Cold Spring Harb Symp Quant Biol 57: 631-642 [Abstract]  
• Hibbs, M., Xu, H, Stacker, S., Springer, T. (1991). Regulation of adhesion of ICAM-1 by the cytoplasmic domain of LFA-1 integrin beta subunit. Science 251: 1611-1613 [Abstract]  
• Uitto, V.-J., Larjava, H. (1991). Extracellular Matrix Molecules and their Receptors: An Overview with Special Emphasis on Periodontal Tissues. CROBM 2: 323-354 [Abstract] [Full Text]  

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