Journal of Cell Science, Vol 107, Issue 3 589-600, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Cell attachment to extracellular matrix substrates is inhibited upon downregulation of expression of calreticulin, an intracellular integrin alpha-subunit-binding protein

CY Leung-Hagesteijn, K Milankov, M Michalak, J Wilkins and S Dedhar
Division of Cancer Research, Sunnybrook Health Science Centre, Toronto, Ontario, Canada.

We have demonstrated recently that calreticulin, an intracellular calcium-binding protein, can interact with the alpha-subunits of integrin receptors via the highly conserved KXGFFKR amino acid sequence present in the cytoplasmic domains of all integrin alpha-subunits (Rojiani et al. (1991) Biochemistry 30, 9859-9866). Here we demonstrate that calreticulin can be co-localized by immunofluorescence as well as co-purified with integrins, that recombinant calreticulin can also interact with integrins, and that the interaction occurs predominantly via the N-domain of calreticulin, to a much lesser extent with the C-domain, but not at all with the proline-rich P-domain. To demonstrate a physiological role for the interaction of calreticulin with integrins, calreticulin expression was downregulated by treating cells with antisense oligonucleotides designed to inhibit the initiation of translation of calreticulin. Antisense oligonucleotides, but not sense or non-sense oligonucleotides, inhibited attachment and spreading of cells cultured in the presence of fetal bovine serum, and also of cells plated on individual extracellular matrix substrates in the absence of serum. The antisense oligonucleotide inhibited cell proliferation of anchorage-dependent cells slightly, but there was no effect on cell viability. The effect on cell attachment was similar to that achieved by treating cells with an antisense oligonucleotide designed to inhibit translation of the integrin alpha 3 subunit, which resulted in the inhibition of cell attachment to alpha 3 beta 1-specific substrates. The effect of the antisense calreticulin oligonucleotide on cell attachment was demonstrated to be integrin-mediated since antisense calreticulin treatment of Jurkat cells abrogated the stimulation of collagen cell attachment achieved by attachment-stimulating signalling anti-alpha 2 (JBS2) and anti-beta 1 (21C8) antibodies. The oligonucleotides did not affect the rate of cell proliferation of these cells. These results demonstrate a fundamental role of calreticulin in cell-extracellular matrix interactions.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. Sipione, C. Ewen, I. Shostak, M. Michalak, and R. C. Bleackley Impaired Cytolytic Activity in Calreticulin-Deficient CTLs J. Immunol., March 15, 2005; 174(6): 3212 - 3219. [Abstract] [Full Text] [PDF]

				C. Bokel, A. Prokop, and N. H. Brown Papillote and Piopio: Drosophila ZP-domain proteins required for cell adhesion to the apical extracellular matrix and microtubule organization J. Cell Sci., February 1, 2005; 118(3): 633 - 642. [Abstract] [Full Text] [PDF]

				C. G. Levine, D. Mitra, A. Sharma, C. L. Smith, and R. S. Hegde The Efficiency of Protein Compartmentalization into the Secretory Pathway Mol. Biol. Cell, January 1, 2005; 16(1): 279 - 291. [Abstract] [Full Text] [PDF]

				S. K. Kang, K. S. Yi, N. S. Kwon, K.-H. Park, U.-H. Kim, K. J. Baek, and M.-J. Im {alpha}1B-Adrenoceptor Signaling and Cell Motility: GTPase FUNCTION OF Gh/TRANSGLUTAMINASE 2 INHIBITS CELL MIGRATION THROUGH INTERACTION WITH CYTOPLASMIC TAIL OF INTEGRIN {alpha} SUBUNITS J. Biol. Chem., August 27, 2004; 279(35): 36593 - 36600. [Abstract] [Full Text] [PDF]

				M. P. Fadel, E. Dziak, C.-M. Lo, J. Ferrier, N. Mesaeli, M. Michalak, and M. Opas Calreticulin Affects Focal Contact-dependent but Not Close Contact-dependent Cell-substratum Adhesion J. Biol. Chem., May 21, 1999; 274(21): 15085 - 15094. [Abstract] [Full Text] [PDF]

				G. Stephens, N. O'Luanaigh, D. Reilly, P. Harriott, B. Walker, D. Fitzgerald, and N. Moran A Sequence within the Cytoplasmic Tail of GpIIb Independently Activates Platelet Aggregation and Thromboxane Synthesis J. Biol. Chem., August 7, 1998; 273(32): 20317 - 20322. [Abstract] [Full Text] [PDF]

				M. Eigenthaler, L. Hofferer, S. J. Shattil, and M. H. Ginsberg A Conserved Sequence Motif in the Integrin beta 3 Cytoplasmic Domain Is Required for Its Specific Interaction with beta 3-Endonexin J. Biol. Chem., March 21, 1997; 272(12): 7693 - 7698. [Abstract] [Full Text] [PDF]

				U. P. Naik, P. M. Patel, and L. V. Parise Identification of a Novel Calcium-binding Protein That Interacts with the Integrin alpha IIb Cytoplasmic Domain J. Biol. Chem., February 21, 1997; 272(8): 4651 - 4654. [Abstract] [Full Text] [PDF]

				M. Michalak, K. Burns, C. Andrin, N. Mesaeli, G. H. Jass, J. L. Busaan, and M. Opas Endoplasmic Reticulum Form of Calreticulin Modulates Glucocorticoid-sensitive Gene Expression J. Biol. Chem., November 15, 1996; 271(46): 29436 - 29445. [Abstract] [Full Text] [PDF]

				E. Chaves-Olarte, I. Florin, P. Boquet, M. Popoff, C. von Eichel-Streiber, and M. Thelestam UDP-Glucose Deficiency in a Mutant Cell Line Protects against Glucosyltransferase Toxins from Clostridium difficile and Clostridium sordellii J. Biol. Chem., March 22, 1996; 271(12): 6925 - 6932. [Abstract] [Full Text] [PDF]

				A. J. Gray, P. W. Park, T. J. Broekelmann, G. J. Laurent, J. T. Reeves, K. R. Stenmark, and R. P. Mecham The Mitogenic Effects of the Bbeta Chain of Fibrinogen Are Mediated through Cell Surface Calreticulin J. Biol. Chem., November 3, 1995; 270(44): 26602 - 26606. [Abstract] [Full Text] [PDF]

				M. Coppolino, C. Leung-Hagesteijn, S. Dedhar, and J. Wilkins Inducible Interaction of Integrin alpha(2)beta(1) with Calreticulin J. Biol. Chem., September 29, 1995; 270(39): 23132 - 23138. [Abstract] [Full Text] [PDF]

				T. K. White, Q. Zhu, and M. L. Tanzer Cell Surface Calreticulin Is a Putative Mannoside Lectin Which Triggers Mouse Melanoma Cell Spreading J. Biol. Chem., July 7, 1995; 270(27): 15926 - 15929. [Abstract] [Full Text] [PDF]

				M. Jannatipour and L. A. Rokeach The Schizosaccharomyces pombe Homologue of the Chaperone Calnexin Is Essential for Viability J. Biol. Chem., March 3, 1995; 270(9): 4845 - 4853. [Abstract] [Full Text] [PDF]

				W. Xu, F. J. Longo, M. R. Wintermantel, X. Jiang, R. A. Clark, and S. DeLisle Calreticulin Modulates Capacitative Ca2+ Influx by Controlling the Extent of Inositol 1,4,5-Trisphosphate-induced Ca2+ Store Depletion J. Biol. Chem., November 17, 2000; 275(47): 36676 - 36682. [Abstract] [Full Text] [PDF]

				S. Goicoechea, A. W. Orr, M. A. Pallero, P. Eggleton, and J. E. Murphy-Ullrich Thrombospondin Mediates Focal Adhesion Disassembly through Interactions with Cell Surface Calreticulin J. Biol. Chem., November 10, 2000; 275(46): 36358 - 36368. [Abstract] [Full Text] [PDF]

				Y. Okazaki, H. Ohno, K. Takase, T. Ochiai, and T. Saito Cell Surface Expression of Calnexin, a Molecular Chaperone in the Endoplasmic Reticulum J. Biol. Chem., November 10, 2000; 275(46): 35751 - 35758. [Abstract] [Full Text] [PDF]

				P. A. Klekotka, S. A. Santoro, H. Wang, and M. M. Zutter Specific Residues within the alpha 2 Integrin Subunit Cytoplasmic Domain Regulate Migration and Cell Cycle Progression via Distinct MAPK Pathways J. Biol. Chem., August 17, 2001; 276(34): 32353 - 32361. [Abstract] [Full Text] [PDF]