HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 29, 30588-30599, July 16, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/29/30588    most recent M313244200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Boutahar, N. Articles by Lafage-Proust, M.-H. Search for Related Content PubMed PubMed Citation Articles by Boutahar, N. Articles by Lafage-Proust, M.-H. Social Bookmarking                      What's this?

Mechanical Strain on Osteoblasts Activates Autophosphorylation of Focal Adhesion Kinase and Proline-rich Tyrosine Kinase 2 Tyrosine Sites Involved in ERK Activation^*

Nadia Boutahar, Alain Guignandon, Laurence Vico, and Marie-Hélène Lafage-Proust

From the Laboratoire de Biologie du Tissu Osseux, INSERM, E366, 15 Rue Ambroise Paré, 42023 Saint-Etienne Cedex 02, France

The mechanisms involved in the mechanical loading-induced increase in bone formation remain unclear. In this study, we showed that cyclic strain (CS) (10 min, 1% stretch at 0.25 Hz) stimulated the proliferation of overnight serum-starved ROS 17/2.8 osteoblast-like cells plated on type I collagen-coated silicone membranes. This increase was blocked by MEK inhibitor PD-98059. Signaling events were then assessed 0 min, 30 min, and 4 h after one CS period with Western blotting and coimmunoprecipitation. CS rapidly and time-dependently promoted phosphorylation of both ERK2 at Tyr-187 and focal adhesion kinase (FAK) at Tyr-397 and Tyr-925, leading to the activation of the Ras/Raf/MEK pathway. Cell transfection with FAK mutated at Tyr-397 completely blocked ERK2 Tyr-187 phosphorylation. Quantitative immunofluorescence analysis of phosphotyrosine residues showed an increase in focal adhesion plaque number and size in strained cells. CS also induced both Src-Tyr-418 phosphorylation and Src to FAK association. Treatment with the selective Src family kinase inhibitor pyrazolopyrimidine 2 did not prevent CS-induced FAK-Tyr-397 phosphorylation suggesting a Src-independent activation of FAK. CS also activated proline-rich tyrosine kinase 2 (PYK2), a tyrosine kinase highly homologous to FAK, at the 402 phosphorylation site and promoted its association to FAK in a time-dependent manner. Mutation of PYK2 at the Tyr-402 site prevented the ERK2 phosphorylation only at 4 h. Intra and extracellular calcium chelators prevented PYK2 activation only at 4 h. In summary, our data showed that osteoblast response to mitogenic CS was mediated by MEK pathway activation. The latter was induced by ERK2 phosphorylation under the control of FAK and PYK2 phosphorylation orchestrated in a time-dependent manner.

Received for publication, December 4, 2003 , and in revised form, March 1, 2004.

^* This work was supported by European Research In Space and Terrestrial Osteoporosis (ERISTO). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 33-477-42-14-54; Fax: 33-477-57-55-72; E-mail: lafagemh{at}univ-st-etienne.fr.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				P. Liu, C. Zhang, J. B. Feng, Y. X. Zhao, X. P. Wang, J. M. Yang, M. X. Zhang, X. L. Wang, and Y. Zhang Cross Talk Among Smad, MAPK, and Integrin Signaling Pathways Enhances Adventitial Fibroblast Functions Activated by Transforming Growth Factor-{beta}1 and Inhibited by Gax Arterioscler. Thromb. Vasc. Biol., April 1, 2008; 28(4): 725 - 731. [Abstract] [Full Text] [PDF]

				C. Dufour, X. Holy, and P. J. Marie Transforming growth factor-{beta} prevents osteoblast apoptosis induced by skeletal unloading via PI3K/Akt, Bcl-2, and phospho-Bad signaling Am J Physiol Endocrinol Metab, April 1, 2008; 294(4): E794 - E801. [Abstract] [Full Text] [PDF]

				Y. Khan and C. T. Laurencin Fracture Repair with Ultrasound: Clinical and Cell-Based Evaluation J. Bone Joint Surg. Am., February 1, 2008; 90(Supplement_1): 138 - 144. [Abstract] [Full Text] [PDF]

				T. Hayashida, M.-H. Wu, A. Pierce, A.-C. Poncelet, J. Varga, and H. W. Schnaper MAP-kinase activity necessary for TGFbeta1-stimulated mesangial cell type I collagen expression requires adhesion-dependent phosphorylation of FAK tyrosine 397 J. Cell Sci., December 1, 2007; 120(23): 4230 - 4240. [Abstract] [Full Text] [PDF]

				Y. Hayashi, M. K. Furue, T. Okamoto, K. Ohnuma, Y. Myoishi, Y. Fukuhara, T. Abe, J. D. Sato, R.-I. Hata, and M. Asashima Integrins Regulate Mouse Embryonic Stem Cell Self-Renewal Stem Cells, December 1, 2007; 25(12): 3005 - 3015. [Abstract] [Full Text] [PDF]

				L. Buckbinder, D. T. Crawford, H. Qi, H. Z. Ke, L. M. Olson, K. R. Long, P. C. Bonnette, A. P. Baumann, J. E. Hambor, W. A. Grasser III, et al. Proline-rich tyrosine kinase 2 regulates osteoprogenitor cells and bone formation, and offers an anabolic treatment approach for osteoporosis PNAS, June 19, 2007; 104(25): 10619 - 10624. [Abstract] [Full Text] [PDF]

				V. David, A. Martin, M.-H. Lafage-Proust, L. Malaval, S. Peyroche, D. B. Jones, L. Vico, and A. Guignandon Mechanical Loading Down-Regulates Peroxisome Proliferator-Activated Receptor {gamma} in Bone Marrow Stromal Cells and Favors Osteoblastogenesis at the Expense of Adipogenesis Endocrinology, May 1, 2007; 148(5): 2553 - 2562. [Abstract] [Full Text] [PDF]

				L. S. Chaturvedi, H. M. Marsh, and M. D. Basson Src and focal adhesion kinase mediate mechanical strain-induced proliferation and ERK1/2 phosphorylation in human H441 pulmonary epithelial cells Am J Physiol Cell Physiol, May 1, 2007; 292(5): C1701 - C1713. [Abstract] [Full Text] [PDF]

				Y. KUMEI, H. SHIMOKAWA, K. OHYA, H. KATANO, H. AKIYAMA, M. HIRANO, and S. MORITA Small GTPase Ras and Rho Expression in Rat Osteoblasts during Spaceflight Ann. N.Y. Acad. Sci., January 1, 2007; 1095(1): 292 - 299. [Abstract] [Full Text] [PDF]

				M. H. Ali, P. T. Mungai, and P. T. Schumacker Stretch-induced phosphorylation of focal adhesion kinase in endothelial cells: role of mitochondrial oxidants Am J Physiol Lung Cell Mol Physiol, July 1, 2006; 291(1): L38 - L45. [Abstract] [Full Text] [PDF]

				J. Rubin, T. C. Murphy, J. Rahnert, H. Song, M. S. Nanes, E. M. Greenfield, H. Jo, and X. Fan Mechanical Inhibition of RANKL Expression Is Regulated by H-Ras-GTPase J. Biol. Chem., January 20, 2006; 281(3): 1412 - 1418. [Abstract] [Full Text] [PDF]

				C. Kesavan, S. Mohan, S. Oberholtzer, J. E. Wergedal, and D. J. Baylink Mechanical loading-induced gene expression and BMD changes are different in two inbred mouse strains J Appl Physiol, November 1, 2005; 99(5): 1951 - 1957. [Abstract] [Full Text] [PDF]

				L. I. Plotkin, I. Mathov, J. I. Aguirre, A. M. Parfitt, S. C. Manolagas, and T. Bellido Mechanical stimulation prevents osteocyte apoptosis: requirement of integrins, Src kinases, and ERKs Am J Physiol Cell Physiol, September 1, 2005; 289(3): C633 - C643. [Abstract] [Full Text] [PDF]

				M. Hughes-Fulford Signal Transduction and Mechanical Stress Sci. Signal., September 7, 2004; 2004(249): re12 - re12. [Abstract] [Full Text] [PDF]

				E. Seeman and G. J. Strewler Clinical and Basic Research Papers - July 2004 Selections IBMS BoneKEy, September 1, 2004; 1(9): 1 - 5. [Full Text] [PDF]