Cell-type specific adhesive interactions of skeletal myoblasts with thrombospondin-1

JC Adams and J Lawler

Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.

Thrombospondin-1 (TSP-1) is an extracellular matrix glycoprotein that may play important roles in the morphogenesis and repair of skeletal muscle. To begin to explore the role of thrombospondin-1 in this tissue, we have examined the interactions of three rodent skeletal muscle cell lines, C2C12, G8, and H9c2, with platelet TSP-1. The cells secrete thrombospondin and incorporate it into the cell layer in a distribution distinct from that of fibronectin. Myoblasts attach and spread on fibronectin- or thrombospondin-coated substrates with similar time and concentration dependencies. Whereas cells adherent on fibronectin organize actin stress fibers, cells adherent on TSP-1 display prominent membrane ruffles and lamellae that contain radial actin microspikes. Attachment to thrombospondin-1 or the 140-kDa tryptic fragment is mediated by interactions with the type 1 repeats and the carboxy-terminal globular domain. Attachment is not inhibited by heparin, GRGDSP peptide, or VTCG peptide but is inhibited by chondroitin sulphate A. Integrins of the beta 1 or alpha V subgroups do not appear to be involved in myoblast attachment to TSP-1; instead, this process depends in part on cell surface chondroitin sulphate proteoglycans. Whereas the central 70-kDa chymotryptic fragment of TSP- 1 does not support myoblast attachment, the carboxy-terminal domain of TSP-1 expressed as a fusion protein in the bacterial expression vector, pGEX, supported myoblast attachment to 30% the level of intact TSP-1. Thrombospondin-4 (TSP-4) is also present in skeletal muscle and a fusion protein containing the carboxy-terminal domain of TSP-4 also supported myoblast adhesion, although this protein was less active on a molar basis than the TSP-1 fusion protein. Thus, the carboxyterminal domain of TSP-1 appears to contain a primary attachment site for myoblasts, and this activity is present in a second member of the thrombospondin family.

Volume 5, Issue 4, pp. 423-437, 04/01/1994
Copyright © 1994 by The American Society for Cell Biology

This article has been cited by other articles:

				R. L. Gundry, K. Raginski, Y. Tarasova, I. Tchernyshyov, D. Bausch-Fluck, S. T. Elliott, K. R. Boheler, J. E. Van Eyk, and B. Wollscheid The Mouse C2C12 Myoblast Cell Surface N-Linked Glycoproteome: IDENTIFICATION, GLYCOSITE OCCUPANCY, AND MEMBRANE ORIENTATION Mol. Cell. Proteomics, November 1, 2009; 8(11): 2555 - 2569. [Abstract] [Full Text] [PDF]

				J. C. Adams, A. A. Bentley, M. Kvansakul, D. Hatherley, and E. Hohenester Extracellular matrix retention of thrombospondin 1 is controlled by its conserved C-terminal region J. Cell Sci., March 15, 2008; 121(6): 784 - 795. [Abstract] [Full Text] [PDF]

				J. I. Zwicker, F. Peyvandi, R. Palla, R. Lombardi, M. T. Canciani, A. Cairo, D. Ardissino, L. Bernardinelli, K. A. Bauer, J. Lawler, et al. The thrombospondin-1 N700S polymorphism is associated with early myocardial infarction without altering von Willebrand factor multimer size Blood, August 15, 2006; 108(4): 1280 - 1283. [Abstract] [Full Text] [PDF]

				T. M. Misenheimer and D. F. Mosher Biophysical Characterization of the Signature Domains of Thrombospondin-4 and Thrombospondin-2 J. Biol. Chem., December 16, 2005; 280(50): 41229 - 41235. [Abstract] [Full Text] [PDF]

				E. Pluskota, O. I. Stenina, I. Krukovets, D. Szpak, E. J. Topol, and E. F. Plow Mechanism and effect of thrombospondin-4 polymorphisms on neutrophil function Blood, December 1, 2005; 106(12): 3970 - 3978. [Abstract] [Full Text] [PDF]

				A. Saumet, M. B. Slimane, M. Lanotte, J. Lawler, and V. Dubernard Type 3 repeat/C-terminal domain of thrombospondin-1 triggers caspase-independent cell death through CD47/{alpha}v{beta}3 in promyelocytic leukemia NB4 cells Blood, July 15, 2005; 106(2): 658 - 667. [Abstract] [Full Text] [PDF]

				E. J. Stevenson, A. Koncarevic, P. G. Giresi, R. W. Jackman, and S. C. Kandarian Transcriptional profile of a myotube starvation model of atrophy J Appl Physiol, April 1, 2005; 98(4): 1396 - 1406. [Abstract] [Full Text] [PDF]

				G. D. Young and J. E. Murphy-Ullrich The Tryptophan-rich Motifs of the Thrombospondin Type 1 Repeats Bind VLAL Motifs in the Latent Transforming Growth Factor-{beta} Complex J. Biol. Chem., November 12, 2004; 279(46): 47633 - 47642. [Abstract] [Full Text] [PDF]

				N. Anilkumar, D. S. Annis, D. F. Mosher, and J. C. Adams Trimeric assembly of the C-terminal region of Thrombospondin-1 or Thrombospondin-2 is necessary for cell spreading and fascin spike organisation J. Cell Sci., January 6, 2002; 115(11): 2357 - 2366. [Abstract] [Full Text] [PDF]

				W.-M. Miao, W. Lin Seng, M. Duquette, P. Lawler, C. Laus, and J. Lawler Thrombospondin-1 Type 1 Repeat Recombinant Proteins Inhibit Tumor Growth through Transforming Growth Factor-{beta}-dependent and -independent Mechanisms Cancer Res., November 1, 2001; 61(21): 7830 - 7839. [Abstract] [Full Text] [PDF]

				R. Riessen, M. Fenchel, H. Chen, D. I. Axel, K. R. Karsch, and J. Lawler Cartilage Oligomeric Matrix Protein (Thrombospondin-5) Is Expressed by Human Vascular Smooth Muscle Cells Arterioscler Thromb Vasc Biol, January 1, 2001; 21(1): 47 - 54. [Abstract] [Full Text] [PDF]

				M. L. Iruela-Arispe, M. Lombardo, H. C. Krutzsch, J. Lawler, and D. D. Roberts Inhibition of Angiogenesis by Thrombospondin-1 Is Mediated by 2 Independent Regions Within the Type 1 Repeats Circulation, September 28, 1999; 100(13): 1423 - 1431. [Abstract] [Full Text] [PDF]

				C. A. Hillery, J. P. Scott, and M. C. Du The Carboxy-Terminal Cell-Binding Domain of Thrombospondin Is Essential for Sickle Red Blood Cell Adhesion Blood, July 1, 1999; 94(1): 302 - 309. [Abstract] [Full Text] [PDF]

				K. Suzuma, H. Takagi, A. Otani, H. Oh, and Y. Honda Expression of Thrombospondin-1 in Ischemia-Induced Retinal Neovascularization Am. J. Pathol., February 1, 1999; 154(2): 343 - 354. [Abstract] [Full Text] [PDF]

				G. Kuznetsov and S. K. Nigam Folding of Secretory and Membrane Proteins N. Engl. J. Med., December 3, 1998; 339(23): 1688 - 1695. [Full Text] [PDF]

				O. V. Volpert, J. Lawler, and N. P. Bouck A human fibrosarcoma inhibits systemic angiogenesis and the growth of experimental metastases via thrombospondin-1 PNAS, May 26, 1998; 95(11): 6343 - 6348. [Abstract] [Full Text] [PDF]

				A. Baumeister, S. Arber, and P. Caroni Accumulation of Muscle Ankyrin Repeat Protein Transcript Reveals Local Activation of Primary Myotube Endcompartments during Muscle Morphogenesis J. Cell Biol., December 1, 1997; 139(5): 1231 - 1242. [Abstract] [Full Text] [PDF]

				J. C. Adams Characterization of Cell-Matrix Adhesion Requirements for the Formation of Fascin Microspikes Mol. Biol. Cell, November 1, 1997; 8(11): 2345 - 2363. [Abstract] [Full Text]

				D. Fischer, R. P. Tucker, R. Chiquet-Ehrismann, and J. C. Adams Cell-Adhesive Responses to Tenascin-C Splice Variants Involve Formation of Fascin Microspikes Mol. Biol. Cell, October 1, 1997; 8(10): 2055 - 2075. [Abstract] [Full Text]

				S. M. Gantt, P. Clavijo, X. Bai, J. D. Esko, and P. Sinnis Cell Adhesion to a Motif Shared by the Malaria Circumsporozoite Protein and Thrombospondin Is Mediated by Its Glycosaminoglycan-binding Region and Not by CSVTCG J. Biol. Chem., August 1, 1997; 272(31): 19205 - 19213. [Abstract] [Full Text] [PDF]

				G. Kuznetsov, L. B. Chen, and S. K. Nigam Multiple Molecular Chaperones Complex with Misfolded Large Oligomeric Glycoproteins in the Endoplasmic Reticulum J. Biol. Chem., January 31, 1997; 272(5): 3057 - 3063. [Abstract] [Full Text] [PDF]

				H. Chen, D. K. Strickland, and D. F. Mosher Metabolism of Thrombospondin 2. BINDING AND DEGRADATION BY 3T3 CELLS AND GLYCOSAMINOGLYCAN-VARIANT CHINESE HAMSTER OVARY CELLS J. Biol. Chem., July 5, 1996; 271(27): 15993 - 15999. [Abstract] [Full Text] [PDF]

				J. Murphy-Ullrich, M. Pallero, N Boerth, J. Greenwood, T. Lincoln, and T. Cornwell Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly J. Cell Sci., January 10, 1996; 109(10): 2499 - 2508. [Abstract] [PDF]

				J. Adams Formation of stable microspikes containing actin and the 55 kDa actin bundling protein, fascin, is a consequence of cell adhesion to thrombospondin-1: implications for the anti-adhesive activities of thrombospondin-1 J. Cell Sci., January 5, 1995; 108(5): 1977 - 1990. [Abstract] [PDF]

				J. Hofsteenge, K. G. Huwiler, B. Macek, D. Hess, J. Lawler, D. F. Mosher, and J. Peter-Katalinic C-Mannosylation and O-Fucosylation of the Thrombospondin Type 1 Module J. Biol. Chem., February 23, 2001; 276(9): 6485 - 6498. [Abstract] [Full Text] [PDF]