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Vol. 10, Issue 11, 3801-3813, November 1999

Roles of Bone Morphogenetic Protein Type I Receptors and Smad Proteins in Osteoblast and Chondroblast Differentiation

Makiko Fujii,*dagger Kohsuke Takeda,*Dagger Takeshi Imamura,* Hiromasa Aoki,* T. Kuber Sampath,§ Shoji Enomoto,parallel Masahiro Kawabata,* Mitsuyasu Kato,* Hidenori Ichijo,*Dagger and Kohei Miyazono*

 *Department of Biochemistry, The Cancer Institute of the Japanese Foundation for Cancer Research and Research for the Future Program, Japan Society for the Promotion of Science, Tokyo 170-8455, Japan;  Dagger Department of Biomaterials Science and  parallel Second Department of Oral Surgery, Faculty of Dentistry, Tokyo Medical and Dental University, Tokyo 113-8549, Japan; and  §Creative BioMolecules, Hopkinton, Massachusetts 01748

The biological effects of type I serine/threonine kinase receptors and Smad proteins were examined using an adenovirus-based vector system. Constitutively active forms of bone morphogenetic protein (BMP) type I receptors (BMPR-IA and BMPR-IB; BMPR-I group) and those of activin receptor-like kinase (ALK)-1 and ALK-2 (ALK-1 group) induced alkaline phosphatase activity in C2C12 cells. Receptor-regulated Smads (R-Smads) that act in the BMP pathways, such as Smad1 and Smad5, also induced the alkaline phosphatase activity in C2C12 cells. BMP-6 dramatically enhanced alkaline phosphatase activity induced by Smad1 or Smad5, probably because of the nuclear translocation of R-Smads triggered by the ligand. Inhibitory Smads, i.e., Smad6 and Smad7, repressed the alkaline phosphatase activity induced by BMP-6 or the type I receptors. Chondrogenic differentiation of ATDC5 cells was induced by the receptors of the BMPR-I group but not by those of the ALK-1 group. However, kinase-inactive forms of the receptors of the ALK-1 and BMPR-I groups blocked chondrogenic differentiation. Although R-Smads failed to induce cartilage nodule formation, inhibitory Smads blocked it. Osteoblast differentiation induced by BMPs is thus mediated mainly via the Smad-signaling pathway, whereas chondrogenic differentiation may be transmitted by Smad-dependent and independent pathways.


dagger    Corresponding author. E-mail address: fjios2{at}dent.tmd.ac.jp.


Molecular Biology of the Cell
Vol. 10, 3801-3813, November 1999
Copyright © 1999 by The American Society for Cell Biology



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J. Skillington, L. Choy, and R. Derynck
Bone morphogenetic protein and retinoic acid signaling cooperate to induce osteoblast differentiation of preadipocytes
J. Cell Biol., October 14, 2002; 159(1): 135 - 146.
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J. Biol. Chem.Home page
U. Valcourt, J. Gouttenoire, A. Moustakas, D. Herbage, and F. Mallein-Gerin
Functions of Transforming Growth Factor-beta Family Type I Receptors and Smad Proteins in the Hypertrophic Maturation and Osteoblastic Differentiation of Chondrocytes
J. Biol. Chem., September 6, 2002; 277(37): 33545 - 33558.
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Mol. Biol. CellHome page
A. Nishihara, T. Watabe, T. Imamura, and K. Miyazono
Functional Heterogeneity of Bone Morphogenetic Protein Receptor-II Mutants Found in Patients with Primary Pulmonary Hypertension
Mol. Biol. Cell, September 1, 2002; 13(9): 3055 - 3063.
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J. Biol. Chem.Home page
C.-F. Lai and S.-L. Cheng
Signal Transductions Induced by Bone Morphogenetic Protein-2 and Transforming Growth Factor-beta in Normal Human Osteoblastic Cells
J. Biol. Chem., May 3, 2002; 277(18): 15514 - 15522.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
B.-C. Kim, M. Mamura, K. S. Choi, B. Calabretta, and S.-J. Kim
Transforming Growth Factor {beta}1 Induces Apoptosis through Cleavage of BAD in a Smad3-Dependent Mechanism in FaO Hepatoma Cells
Mol. Cell. Biol., March 1, 2002; 22(5): 1369 - 1378.
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J. Cell Sci.Home page
A. Hoffmann, S. Czichos, C. Kaps, D. Bachner, H. Mayer, Y. Zilberman, G. Turgeman, G. Pelled, G. Gross, and D. Gazit
The T-box transcription factor Brachyury mediates cartilage development in mesenchymal stem cell line C3H10T1/2
J. Cell Sci., February 15, 2002; 115(4): 769 - 781.
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J. Biol. Chem.Home page
S. Bai and X. Cao
A Nuclear Antagonistic Mechanism of Inhibitory Smads in Transforming Growth Factor-beta Signaling
J. Biol. Chem., February 1, 2002; 277(6): 4176 - 4182.
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J. Biol. Chem.Home page
W. Samuel, C. N. Nagineni, R. K. Kutty, W. T. Parks, J. S. Gordon, S. M. Prouty, J. J. Hooks, and B. Wiggert
Transforming Growth Factor-beta Regulates Stearoyl Coenzyme A Desaturase Expression through a Smad Signaling Pathway
J. Biol. Chem., January 4, 2002; 277(1): 59 - 66.
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Genome ResHome page
J. Theilhaber, T. Connolly, S. Roman-Roman, S. Bushnell, A. Jackson, K. Call, T. Garcia, and R. Baron
Finding Genes in the C2C12 Osteogenic Pathway by k-Nearest-Neighbor Classification of Expression Data
Genome Res., January 1, 2002; 12(1): 165 - 176.
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JCBHome page
A. Hanyu, Y. Ishidou, T. Ebisawa, T. Shimanuki, T. Imamura, and K. Miyazono
The N domain of Smad7 is essential for specific inhibition of transforming growth factor-{beta} signaling
J. Cell Biol., December 10, 2001; 155(6): 1017 - 1028.
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J. Biol. Chem.Home page
P. Tylzanowski, K. Verschueren, D. Huylebroeck, and F. P. Luyten
Smad-interacting Protein 1 Is a Repressor of Liver/Bone/Kidney Alkaline Phosphatase Transcription in Bone Morphogenetic Protein-induced Osteogenic Differentiation of C2C12 Cells
J. Biol. Chem., October 19, 2001; 276(43): 40001 - 40007.
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J. Cell Sci.Home page
H Aoki, M Fujii, T Imamura, K Yagi, K Takehara, M Kato, and K Miyazono
Synergistic effects of different bone morphogenetic protein type I receptors on alkaline phosphatase induction
J. Cell Sci., January 4, 2001; 114(8): 1483 - 1489.
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EndocrinologyHome page
F. De Luca, K. M. Barnes, J. A. Uyeda, S. De-Levi, V. Abad, T. Palese, V. Mericq, and J. Baron
Regulation of Growth Plate Chondrogenesis by Bone Morphogenetic Protein-2
Endocrinology, January 1, 2001; 142(1): 430 - 436.
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Mol. Cell. Biol.Home page
K.-S. Lee, H.-J. Kim, Q.-L. Li, X.-Z. Chi, C. Ueta, T. Komori, J. M. Wozney, E.-G. Kim, J.-Y. Choi, H.-M. Ryoo, et al.
Runx2 Is a Common Target of Transforming Growth Factor beta 1 and Bone Morphogenetic Protein 2, and Cooperation between Runx2 and Smad5 Induces Osteoblast-Specific Gene Expression in the Pluripotent Mesenchymal Precursor Cell Line C2C12
Mol. Cell. Biol., December 1, 2000; 20(23): 8783 - 8792.
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Proc. Natl. Acad. Sci. USAHome page
Y.-W. Zhang, N. Yasui, K. Ito, G. Huang, M. Fujii, J.-i. Hanai, H. Nogami, T. Ochi, K. Miyazono, and Y. Ito
A RUNX2/PEBP2alpha A/CBFA1 mutation displaying impaired transactivation and Smad interaction in cleidocranial dysplasia
PNAS, August 23, 2000; (2000) 180309597.
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Endocr. Rev.Home page
A. Yamaguchi, T. Komori, and T. Suda
Regulation of Osteoblast Differentiation Mediated by Bone Morphogenetic Proteins, Hedgehogs, and Cbfa1
Endocr. Rev., August 1, 2000; 21(4): 393 - 411.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
W. Ishida, T. Hamamoto, K. Kusanagi, K. Yagi, M. Kawabata, K. Takehara, T. K. Sampath, M. Kato, and K. Miyazono
Smad6 Is a Smad1/5-induced Smad Inhibitor. CHARACTERIZATION OF BONE MORPHOGENETIC PROTEIN-RESPONSIVE ELEMENT IN THE MOUSE Smad6 PROMOTER
J. Biol. Chem., February 25, 2000; 275(9): 6075 - 6079.
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J. Biol. Chem.Home page
H. Watanabe, M. P. de Caestecker, and Y. Yamada
Transcriptional Cross-talk between Smad, ERK1/2, and p38 Mitogen-activated Protein Kinase Pathways Regulates Transforming Growth Factor-beta -induced Aggrecan Gene Expression in Chondrogenic ATDC5 Cells
J. Biol. Chem., April 20, 2001; 276(17): 14466 - 14473.
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J. Biol. Chem.Home page
S. Nishimori, Y. Tanaka, T. Chiba, M. Fujii, T. Imamura, K. Miyazono, T. Ogasawara, H. Kawaguchi, T. Igarashi, T. Fujita, et al.
Smad-mediated Transcription Is Required for Transforming Growth Factor-beta 1-induced p57Kip2 Proteolysis in Osteoblastic Cells
J. Biol. Chem., March 30, 2001; 276(14): 10700 - 10705.
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J. Biol. Chem.Home page
M. Izumi, Y. Fujio, K. Kunisada, S. Negoro, E. Tone, M. Funamoto, T. Osugi, Y. Oshima, Y. Nakaoka, T. Kishimoto, et al.
Bone Morphogenetic Protein-2 Inhibits Serum Deprivation-induced Apoptosis of Neonatal Cardiac Myocytes through Activation of the Smad1 Pathway
J. Biol. Chem., August 10, 2001; 276(33): 31133 - 31141.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
E. Piek, W. J. Ju, J. Heyer, D. Escalante-Alcalde, C. L. Stewart, M. Weinstein, C. Deng, R. Kucherlapati, E. P. Bottinger, and A. B. Roberts
Functional Characterization of Transforming Growth Factor beta Signaling in Smad2- and Smad3-deficient Fibroblasts
J. Biol. Chem., June 1, 2001; 276(23): 19945 - 19953.
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J. Biol. Chem.Home page
K. Pardali, A. Kurisaki, A. Moren, P. ten Dijke, D. Kardassis, and A. Moustakas
Role of Smad Proteins and Transcription Factor Sp1 in p21Waf1/Cip1 Regulation by Transforming Growth Factor-beta
J. Biol. Chem., September 15, 2000; 275(38): 29244 - 29256.
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Proc. Natl. Acad. Sci. USAHome page
Y.-W. Zhang, N. Yasui, K. Ito, G. Huang, M. Fujii, J.-i. Hanai, H. Nogami, T. Ochi, K. Miyazono, and Y. Ito
A RUNX2/PEBP2alpha A/CBFA1 mutation displaying impaired transactivation and Smad interaction in cleidocranial dysplasia
PNAS, September 12, 2000; 97(19): 10549 - 10554.
[Abstract] [Full Text] [PDF]




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