Bone Morphogenetic Protein-2 Stimulates Runx2 Acetylation

Abstract

Runx2/Cbfa1/Pebp2aA is a global regulator of osteogenesis and is crucial for regulating the expression of bone-specific genes. Runx2 is a major target of the bone morphogenetic protein (BMP) pathway. Genetic analysis has revealed that Runx2 is degraded through a Smurf-mediated ubiquitination pathway, and its activity is inhibited by HDAC4. Here, we demonstrate the molecular link between Smurf, HDACs and Runx2, in BMP signaling. BMP-2 signaling stimulates p300-mediated Runx2 acetylation, increasing transactivation activity and inhibiting Smurf1-mediated degradation of Runx2. HDAC4 and HDAC5 dea-cetylate Runx2, allowing the protein to undergo Smurf-mediated degradation. Inhibition of HDAC increases Runx2 acetylation, and potentiates BMP-2-stimulated osteoblast differentiation and increases bone formation. These results demonstrate that the level of Runx2 is controlled by a dynamic equilibrium of acetylation, deacetylation, and ubiquitination. These findings have important medical implications because BMPs and Runx2 are of tremendous interest with regard to the development of therapeutic agents against bone diseases. Runx2/Cbfa1/Pebp2aA is a global regulator of osteogenesis and is crucial for regulating the expression of bone-specific genes. Runx2 is a major target of the bone morphogenetic protein (BMP) pathway. Genetic analysis has revealed that Runx2 is degraded through a Smurf-mediated ubiquitination pathway, and its activity is inhibited by HDAC4. Here, we demonstrate the molecular link between Smurf, HDACs and Runx2, in BMP signaling. BMP-2 signaling stimulates p300-mediated Runx2 acetylation, increasing transactivation activity and inhibiting Smurf1-mediated degradation of Runx2. HDAC4 and HDAC5 dea-cetylate Runx2, allowing the protein to undergo Smurf-mediated degradation. Inhibition of HDAC increases Runx2 acetylation, and potentiates BMP-2-stimulated osteoblast differentiation and increases bone formation. These results demonstrate that the level of Runx2 is controlled by a dynamic equilibrium of acetylation, deacetylation, and ubiquitination. These findings have important medical implications because BMPs and Runx2 are of tremendous interest with regard to the development of therapeutic agents against bone diseases. Runx2/Cbfa1/Pebp2αA is a transcription factor belonging to the Runx family, which contains three members: Runx1, Runx2, and Runx3 (1van Wijnen A.J. Stein G.S. Gergen J.P. Groner Y. Hiebert S.W. Ito Y. Liu P. Neil J.C. Ohki M. Speck N. Oncogene. 2004; 23: 4209-4210Crossref PubMed Scopus (99) Google Scholar). The importance of Runx proteins in mammalian development has been clearly demonstrated by gene disruption studies (2Ito Y. Oncogene. 2004; 23: 4198-4208Crossref PubMed Scopus (279) Google Scholar). Mouse embryos with homozygous mutations in Runx1 showed normal morphogenesis and yolk sac-derived primitive hematopoiesis, they lacked fetal liver hematopoiesis, indicating that Runx1 is required for definitive hematopoiesis (3Okuda T. van Deursen J. Hiebert S.W. Grosveld G. Downing J.R. Cell. 1996; 84: 321-330Abstract Full Text Full Text PDF PubMed Scopus (1613) Google Scholar). Runx3 functions as a tumor suppressor of gastric cancer. Runx3-null gastric mucosa develops hyperplasia because of the activation of proliferation and suppression of apoptosis in epithelial cells. Runx3-null gastric epithelial cells are less sensitive to the growth-inhibiting and apoptosis-inducing activities of transforming growth factor-β (TGF-β). 2The abbreviations used are: TGF-β, transforming growth factor-β; HDAC, histone deacetylase; BMP, bone morphogenetic protein-2; OC, osteocalcin; siRNA, small interfering RNA; HA, hemagglutinin; Smurf1, Smad ubiquitin regulatory factor 1; TSA, Tricostatin A; ALP, alkaline phosphatase; RT, reverse transcriptase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; CMV, cytomegalovirus. Thus, Runx3 is a major growth regulator of gastric epithelial cells. Previously, we showed that the extent of the Runx3 acetylation is up-regulated by the TGF-β signaling pathway and down-regulated by histone deacetylase (HDAC) activities (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google Scholar). Runx2 is a key regulator of chondroblast and osteoblast differentiation, and of bone development in vivo (5Ducy P. Zhang R. Geoffroy V. Ridall A.L. Karsenty G. Cell. 1997; 89: 747-754Abstract Full Text Full Text PDF PubMed Scopus (3668) Google Scholar, 6Fujita T. Azuma Y. Fukuyama R. Hattori Y. Yoshida C. Koida M. Ogita K. Komori T. J. Cell Biol. 2004; 166: 85-95Crossref PubMed Scopus (362) Google Scholar, 7Yoshida C.A. Yamamoto H. Fujita T. Furuichi T. Ito K. Inoue K. Yamana K. Zanma A. Takada K. Ito Y. Komori T. Genes Dev. 2004; 18: 952-963Crossref PubMed Scopus (462) Google Scholar). Targeted disruption of this Runx2 in mice resulted in a complete lack of ossification due to the maturational arrest of osteoblasts (8Komori T. Yagi H. Nomura S. Yamaguchi A. Sasaki K. Deguchi K. Shimizu Y. Bronson R.T. Gao Y.H. Inada M. Sato M. Okamoto R. Kitamura Y. Yoshiki S. Kishimoto T. Cell. 1997; 89: 755-764Abstract Full Text Full Text PDF PubMed Scopus (3678) Google Scholar, 9Otto F. Thornell A.P. Crompton T. Denzel A. Gilmour K.C. Rosewell I.R. Stamp G.W. Beddington R.S. Mundlos S. Olsen B.R. Selby P.B. Owen M.J. Cell. 1997; 89: 765-771Abstract Full Text Full Text PDF PubMed Scopus (2430) Google Scholar, 10Choi J.Y. Pratap J. Javed A. Zaidi S.K. Xing L. Balint E. Dalamangas S. Boyce B. van Wijnen A.J. Lian J.B. Stein J.L. Jones S.N. Stein G.S. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 8650-8655Crossref PubMed Scopus (245) Google Scholar). Runx2 regulates the expression of major extracellular matrix genes (including alkaline phosphatase, osteopontin, osteocalcin, type I collagen, and type X collagen) expressed by chondroblasts and osteoblasts (5Ducy P. Zhang R. Geoffroy V. Ridall A.L. Karsenty G. Cell. 1997; 89: 747-754Abstract Full Text Full Text PDF PubMed Scopus (3668) Google Scholar, 11Banerjee C. McCabe L.R. Choi J.Y. Hiebert S.W. Stein J.L. Stein G.S. Lian J.B. J. Cell. Biochem. 1997; 66: 1-8Crossref PubMed Scopus (398) Google Scholar). Inactivation of one allele of the RUNX2 gene was identified as the cause of human cleidocranial dysplasia (CCD), an autosomal dominant bone disorder (12Lee B. Thirunavukkarasu K. Zhou L. Pastore L. Baldini A. Hecht J. Geoffroy V. Ducy P. Karsenty G. Nat. Genet. 1997; 16: 307-310Crossref PubMed Scopus (497) Google Scholar, 13Mundlos S. Otto F. Mundlos C. Mulliken J.B. Aylsworth A.S. Albright S. Lindhout D. Cole W.G. Henn W. Knoll J.H. Owen M.J. Mertelsmann R. Zabel B.U. Olsen B.R. Cell. 1997; 89: 773-779Abstract Full Text Full Text PDF PubMed Scopus (1285) Google Scholar). On the other hand, constitutively active mutation of FGFR1 results in increased Runx2 expression and the premature suture closure that is the hallmark of human craniosynostosis (14Zhou Y.X. Xu X. Chen L. Li C. Brodie S.G. Deng C.X. Hum. Mol. Genet. 2000; 9: 2001-2008Crossref PubMed Scopus (201) Google Scholar). Expression and activation of Runx2 is regulated by many bone-derived growth factors, including bone morphogenetic proteins (BMPs) (5Ducy P. Zhang R. Geoffroy V. Ridall A.L. Karsenty G. Cell. 1997; 89: 747-754Abstract Full Text Full Text PDF PubMed Scopus (3668) Google Scholar, 15Lee K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar). BMPs form a unique group of proteins within the TGF-β superfamily of genes, and play pivotal roles in the regulation of heart, neural, cartilage, and bone development (16Chen D. Zhao M. Mundy G.R. Growth Factors. 2004; 22: PubMed Scopus Google Scholar, J.M. V. A.J. M.J. PubMed Scopus Google Scholar). The Smad of proteins has been identified as the of BMP Y. J. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). and T. Yagi H. Nomura S. Yamaguchi A. Sasaki K. Deguchi K. Shimizu Y. Bronson R.T. Gao Y.H. Inada M. Sato M. Okamoto R. Kitamura Y. Yoshiki S. Kishimoto T. Cell. 1997; 89: 755-764Abstract Full Text Full Text PDF PubMed Scopus (3678) Google Runx2 gene expression K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google and with the Runx2 protein to osteoblast differentiation K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar, J. Chen T. N. Kim T. Y. M. M.J. J. M. K. Ito Y. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, N. Ito K. G. M. J. H. T. K. Ito Y. Proc. Natl. Acad. Sci. U. S. A. 2000; PubMed Scopus Google Scholar). The protein functions as a of transcription for a of proteins including Runx D. J. 1997; PubMed Scopus Google Scholar, C. L. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, V. K. A. A.S. K. Cell. 1997; 89: Full Text Full Text PDF PubMed Scopus Google Scholar, A. Shimizu K. Ohki M. J. PubMed Scopus Google and functions as a histone V. Y. Cell. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). of histone is to the and to a for transcription A.P. D. Cell. 1996; 84: Full Text Full Text PDF PubMed Scopus Google Scholar). is of a of proteins T. J. 2000; PubMed Scopus Google Scholar). is by and the acetylation is by histone indicating that acetylation is a for regulation of proteins G. C. P. K. M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The in through acetylation to the protein acetylation and ubiquitination A. Y. Y. E. J. PubMed Scopus Google Scholar, E. U. J. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). ubiquitination a crucial in and protein degradation by Runx2 is degraded through a pathway G. K. Ito K. H.J. T. Ito Y. J. 2001; 20: PubMed Scopus Google Scholar, Y. F. V. Karsenty G. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). ubiquitin regulatory factor was identified as the ubiquitin for Runx2 degradation (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google Scholar, M. M. Mundy G.R. Chen D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). of in osteoblast cells BMP signaling and osteoblast differentiation M. M. Mundy G.R. Chen D. J. Biol. Chem. 2004; 279: Full Text Full Text PDF PubMed Scopus Google Scholar). with of resulted in bone M. Zhang Li C. Deng C.X. Zhang Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). mice premature ossification of the that results Runx2 a was between and the HDAC4 (8Komori T. Yagi H. Nomura S. Yamaguchi A. Sasaki K. Deguchi K. Shimizu Y. Bronson R.T. Gao Y.H. Inada M. Sato M. Okamoto R. Kitamura Y. Yoshiki S. Kishimoto T. Cell. 1997; 89: 755-764Abstract Full Text Full Text PDF PubMed Scopus (3678) Google Scholar, 9Otto F. Thornell A.P. Crompton T. Denzel A. Gilmour K.C. Rosewell I.R. Stamp G.W. Beddington R.S. Mundlos S. Olsen B.R. Selby P.B. Owen M.J. Cell. 1997; 89: 765-771Abstract Full Text Full Text PDF PubMed Scopus (2430) Google Scholar, M. T. Nomura S. S. Deguchi K. M. Sato M. H. T. N. T. N. Kitamura Y. Kishimoto T. Komori T. Dev. PubMed Scopus Google Scholar, K. J. X. E. J. C. J.M. Karsenty G. Cell. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar, C. M. N. Yoshida C. Liu Y. M. T. H. K. Takada K. K. Komori T. J. Cell Biol. 2001; PubMed Scopus Google Scholar). demonstrated that HDAC4 is required to Runx2 T. R. R. J. PubMed Scopus Google that HDAC4 is a of Runx2. results clearly demonstrate that and HDAC play roles in BMP signaling of Runx2 the molecular have been clearly this we the molecular that HDAC and Runx2. that BMP-2 Runx2 acetylation for the an important molecular link between in Runx2, HDAC, and Smurf1, and one that a of one Cell and and cells in with fetal and in was of Runx2 and in fetal and cells used for the which required BMP signaling pathway. was used of was cells used was required and BMP signaling pathway was and Runx2 type I with and its with HA, in a mammalian expression and the Runx2 acetylation by and the The BMP and expression K. and M. The expression for the The which contains of identified in the M.J. J. J. Google was K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar). The of was the and Javed A. Kim H.J. S. Choi J.Y. V. Stein J.L. van Wijnen A.J. Stein G.S. Lian J.B. Ryoo H.M. J. Cell. Biochem. PubMed Scopus Google Scholar). The and RUNX2 of the have been H.J. Kim J.H. Bae S.C. Choi J.Y. Kim H.J. Ryoo H.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). this against and The was by a with a Runx2 and the for cells for the the to the The was as an to the of and the and cells in and and by The with the and and protein protein The and proteins and to The with the and with the proteins with an HDAC and cells in for and with the Y. K.Y. M. T. N. Sato S. T. T. R. Yoshida M. S. H. 2001; Google and N. B. S. S. T. Y. Yoshida M. PubMed Scopus Google for the in The by M. Yoshida cells for in of of and A. The cells in and the protein was with The was with protein with with and to for to cells cells in and in fetal in the of with a was with and HDAC4 siRNA, and HDAC5 and and was used as a the and to analysis and Mouse of mice and analysis of bone was as by I.R. Mol. Google The of bone was the BMP-2 Runx2 a a to the of osteoblast differentiation bone formation. this BMP-2 the differentiation of and osteoblast differentiation T. Yamaguchi A. M. E. N. T. V. Wozney J.M. A. T. J. Cell Biol. PubMed Scopus Google Scholar). Runx2 is a major target of the BMP-2 signaling pathway the level K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar). BMP-2 regulates Runx2 as cells with Runx2, in the of and the protein by The level of expressed Runx2 was in cells was by of Runx2 and increasing of the BMP Runx2 increased in to the of The of Runx2 was in the of and to in the of and the of BMP-2 activation of the we of and Runx2, BMP-2 increased the level of Runx2 the level was and These results in to BMP-2 the of Runx2 is of the in is with increased Runx2 and Runx2 and Runx3 are degraded the pathway (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google Scholar, G. K. Ito K. H.J. T. Ito Y. J. 2001; 20: PubMed Scopus Google Scholar, Y. F. V. Karsenty G. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and acetylation proteins ubiquitination (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google Scholar, A. Y. Y. E. J. PubMed Scopus Google we BMP-2 Runx2 acetylation, the Runx2 analysis of cells revealed that Runx2 is by BMP-2 is with TGF-β superfamily signaling and Runx1 and Runx3 are for activity (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google Scholar, A. Shimizu K. Ohki M. J. PubMed Scopus Google Scholar, E. U. J. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google we is with Runx2 Runx2, and p300-mediated Runx2 acetylation increased in the of BMP-2 the of Runx2 was to acetylation of in with resulted in an in Runx2 acetylation and an of the protein and of Runx2 was by BMP-2 of Runx2 acetylation by and BMP-2 was in which Runx2 was with of Runx2 increased was and increased in to BMP-2 Runx2 is by cells with expression and the acetylation of Runx2 was of increased Runx2 acetylation and Runx2 These results demonstrate that the BMP-2 signaling pathway stimulates p300-mediated Runx2 acetylation, Runx2 Smad for p300-mediated Runx2 BMP-2 stimulates p300-mediated Runx2 acetylation, we cells with and Runx2 expression and Runx2 and and and the level of Runx2 acetylation and are of BMP-2 we the of and p300-mediated Runx2 of and was by and The level of Runx2 acetylation was by and by and Runx2 acetylation of and increased Runx2 acetylation that Runx2 acetylation in to the of in Runx2 acetylation, and in cells. The between Runx2 and increased the The between the of and Runx2 acetylation and the that p300-mediated Runx2 acetylation by the between Runx2 and Runx2 Smurf1-mediated contains that are acetylation which are for acetylation, a of Runx2 with and an was in the and of Runx2 the Runx2 with for The Runx2 acetylation by this we acetylation is for of the Runx2 a of with the was in the of These results that the of increases the of the protein to and that acetylation of the Runx2 degradation. Runx2 is degraded through the pathway and is the ubiquitin for this pathway M. M. Mundy G.R. Chen D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, M. M. Mundy G.R. Chen D. J. Biol. Chem. 2004; 279: Full Text Full Text PDF PubMed Scopus Google we acetylation Runx2 Smurf1-mediated degradation. Runx2 was degraded is of the Smurf1-mediated Runx2 degradation in the of the was to Smurf1-mediated degradation. Runx2 contains a that is required for with the of L. U. B. J.R. M. P. G. R. J. H. Sci. PubMed Scopus Google Runx2 we the which the and the which has a the the and and to Smurf1-mediated degradation These results are with Smurf1-mediated degradation of Runx2. HDAC has been that Runx3 acetylation is down-regulated by HDACs (4Jin Y.H. Jeon E.J. Li Q.L. Lee Y.H. Choi J.K. Kim W.J. Lee K.Y. Bae S.C. J. Biol. Chem. 2004; 279: 29409-29417Abstract Full Text Full Text PDF PubMed Scopus (184) Google and Runx2 activity is by HDACs T. R. R. J. PubMed Scopus Google and increased by of HDAC4 K. J. X. E. J. C. J.M. Karsenty G. Cell. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). we the of HDAC4 and Runx2. and analysis revealed that HDAC4 and with Runx2, and Runx2 protein the of of the Runx2 we of Runx2 and GAPDH, The level of Runx2 was inhibited that the of Runx2 is of the in is with the protein HDAC4 and p300-mediated Runx2 acetylation, and Runx2 protein with this of HDAC4 resulted in an in Runx2 acetylation, and Runx2 was the HDACs inhibited by Tricostatin and These results demonstrate that Runx2 is a dynamic and that HDAC4 and the of Runx2 Runx2 for Runx2 acetylation its transactivation the TGF-β and Runx2 expression cells. cells with increasing of HDAC The HDAC increased the activity in a These findings that the transactivation activity of Runx2 increased by inhibiting Runx2 in a in the in transactivation activity of Runx2 by acetylation is a of increased protein of the acetylation the transactivation activities of Runx2 and in the of Runx2 increased activity by to and with increased activity by mutation of the by a in Runx2 transactivation activity the of the proteins was increased and to the activity of the Runx2 proteins the of Runx2 with These results that Runx2 acetylation is required for for transactivation HDAC in Runx2 is a key regulator of osteoblast differentiation, and a major target of BMP-2 we HDAC which Runx2 acetylation, osteoblast is an that is in the of osteoblast differentiation T. Yamaguchi A. M. E. N. T. V. Wozney J.M. A. T. J. Cell Biol. PubMed Scopus Google Scholar). of cells with HDAC a in activity The increased of Runx2 BMP-2 activity in osteoblast differentiation K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar). this cells with of BMP-2 with increasing of HDAC BMP-2 of the with HDAC BMP-2 increased activity in a OC, is expressed the osteoblast J.B. Stein G.S. Stein J.L. van Wijnen A.J. J. Cell. Biochem. PubMed Google Scholar). that Runx2 acetylation osteoblast differentiation, we the of HDAC the and the gene the of HDAC Runx2, we Runx2 expression and in which and the cells with the HDAC the of Runx2, the HDAC the Runx2 the in cells. the of Runx2, HDAC the the activation for Runx2 The disruption of a in the by HDAC by Runx2 HDAC gene expression was by and the of HDAC expression with Runx2 gene and this was increased by HDAC and analysis of that HDAC expression increased in the of Runx2 is with the results of activity analysis the in Runx2 acetylation by HDAC osteoblast Expression by Runx2 expression regulated by other transcription factors, as Kim Kim H.J. H.M. J.H. Wozney J.M. Kim H.J. Ryoo H.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google K. Zhou X. G. Zhang Deng J.M. B. Cell. Full Text Full Text PDF PubMed Scopus Google we Runx2 is the target of HDAC for activation of the The of and the of HDAC the of HDAC in of expression expression in cells the of Runx2 and HDAC expression in the These results three transcription factors, Runx2 is the target of HDAC in osteoblast HDAC the of HDAC bone a of bone was used as a G. R. S. J. Chen D. G. Boyce B. Zhao M. G. PubMed Scopus Google Scholar). with The bone increased in the of with that of The of is to G. R. S. J. Chen D. G. Boyce B. Zhao M. G. PubMed Scopus Google Scholar). analysis is in These results that of Runx2 acetylation by HDAC osteoblast differentiation increases bone formation. BMPs play pivotal roles in the regulation of bone and Runx2 has been identified as a gene osteoblast differentiation, and as a major target of BMP-2 signaling Runx2 through transcription of the Runx2 gene K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar). with the Runx2 protein and to osteoblast differentiation K.S. Kim H.J. Li Q.L. Chi X.Z. Ueta C. Komori T. Wozney J.M. Kim E.G. Choi J.Y. Ryoo H.M. Bae S.C. Mol. Cell. Biol. 2000; 20: 8783-8792Crossref PubMed Scopus (763) Google Scholar, J. Chen T. N. Kim T. Y. M. M.J. J. M. K. Ito Y. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, N. Ito K. G. M. J. H. T. K. Ito Y. Proc. Natl. Acad. Sci. U. S. A. 2000; PubMed Scopus Google Scholar). this we demonstrate a BMP-2 signaling in Runx2 Runx2 Smurf1-mediated degradation. Runx2 acetylation is by the activity of and is by in Runx2 cells with BMP-2 and acetylation mutation of the Runx2, inhibited Smurf1-mediated Runx2 degradation in the acetylation of Runx2, of HDAC, osteoblast differentiation, and increased bone and The acetylation of Runx2 was increased by with BMP-2 and was by acetylation was increased by of and was by of These results that Runx2 acetylation is regulated by the BMP-2 signaling pathway, and p300-mediated Runx2 acetylation by is is the of this acetylation, because and Runx2 acetylation that acetylation of is required for osteoblast differentiation J. H. Lian J.B. Stein G.S. Stein J.L. Wijnen A.J. Mol. PubMed Scopus Google Scholar). cells showed acetylation of and osteoblasts showed acetylation is to the that the of osteoblast gene including that of OC, in bone cells J. H. Lian J.B. Stein G.S. Stein J.L. Wijnen A.J. Mol. PubMed Scopus Google Scholar). this we for acetylation in osteoblast the transactivation activity of Runx2, and increased its by Smurf1-mediated degradation. Runx2 to osteoblast results the of a of gene by histone acetylation J. H. Lian J.B. Stein G.S. Stein J.L. Wijnen A.J. Mol. PubMed Scopus Google because expression was increased by HDAC the importance of Runx2 acetylation is by the results of activity in which results that other transcription factors, as and are by acetylation that increased acetylation and are important for Runx2 because mutations of the by the resulted in a in Runx2 transactivation activity that the acetylation of the Smurf1-mediated is required for transactivation The of an group to a a for protein the with proteins with T. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, Nat. Mol. Cell. Biol. PubMed Scopus Google the functions as a for T. J. 2000; PubMed Scopus Google Scholar). an to in the is acetylation of Runx2 a for protein the of the protein to other regulatory Previously, K. J. X. E. J. C. J.M. Karsenty G. Cell. 2004; Full Text Full Text PDF PubMed Scopus Google that HDAC4 Runx2 activity by Runx2 Runx2 acetylation was results clearly a for HDAC4 and deacetylase activity to acetylation, and activity of Runx2. other have that HDAC increased activation of the Li X. J. Biol. Chem. 2004; 279: Full Text Full Text PDF PubMed Scopus Google and osteoblast and differentiation J. 20: PubMed Scopus (201) Google Scholar, Kim Bae J. Cell. Biochem. PubMed Scopus Google Scholar). of the of Runx2. results that differentiation regulation of the Runx2 protein by and and have been to with Runx2 Li X. J. Biol. Chem. 2004; 279: Full Text Full Text PDF PubMed Scopus Google Scholar, Zaidi S.K. R. van Wijnen A.J. Lian J.B. Yoshida M. Stein G.S. Li X. Mol. Cell. Biol. 22: PubMed Scopus Google that are HDACs Runx2. The link between BMP-2 signaling and Runx2 acetylation is by that HDAC Runx2 with HDAC increased Runx2 acetylation BMP-2 activity in of osteoblast differentiation and bone These results that and ubiquitination are the and activity of Runx2, which is for osteoblast of BMPs and Runx2 tremendous for development as therapeutic agents to bone and bone is that are for E. C. J. 2004; PubMed Scopus Google Scholar). the of of and activation of osteoblasts are with that bone is a for osteoblast that bone Mundy G. R. S. J. Chen D. G. Boyce B. Zhao M. G. PubMed Scopus Google an important in the development of osteoblast by that the BMP-2 that and BMP-2 increasing bone G. R. S. J. Chen D. G. Boyce B. Zhao M. G. PubMed Scopus Google Scholar). the we that of the activity of by increasing Runx2 activity and results a for therapeutic agents against