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Molecular and Cellular Biology, December 2006, p. 9456-9470, Vol. 26, No. 24
0270-7306/06/$08.00+0     doi:10.1128/MCB.00759-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Bone Morphogenetic Protein-Induced Msx1 and Msx2 Inhibit Myocardin-Dependent Smooth Muscle Gene Transcription{triangledown}

Ken'ichiro Hayashi,1 Seiji Nakamura,1,2 Wataru Nishida,1,{dagger} and Kenji Sobue1*

Department of Neuroscience (D13), Osaka University Graduate School of Medicine, Yamadaoka 2-2, Suita, Osaka 565-0871,1 Department of Fixed Prosthodontics, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan2

Received 2 May 2006/ Returned for modification 13 June 2006/ Accepted 22 September 2006

During the onset and progression of atherosclerosis, the vascular smooth muscle cell (VSMC) phenotype changes from differentiated to dedifferentiated, and in some cases, this change is accompanied by osteogenic transition, resulting in vascular calcification. One characteristic of dedifferentiated VSMCs is the down-regulation of smooth muscle cell (SMC) marker gene expression. Bone morphogenetic proteins (BMPs), which are involved in the induction of osteogenic gene expression, are detected in calcified vasculature. In this study, we found that the BMP2-, BMP4-, and BMP6-induced expression of Msx transcription factors (Msx1 and Msx2) preceded the down-regulation of SMC marker expression in cultured differentiated VSMCs. Either Msx1 or Msx2 markedly reduced the myocardin-dependent promoter activities of SMC marker genes (SM22{alpha} and caldesmon). We further investigated interactions between Msx1 and myocardin/serum response factor (SRF)/CArG-box motif (cis element for SRF) using coimmunoprecipitation, gel-shift, and chromatin immunoprecipitation assays. Our results showed that Msx1 or Msx2 formed a ternary complex with SRF and myocardin and inhibited the binding of SRF or SRF/myocardin to the CArG-box motif, resulting in inhibition of their transcription.

* Corresponding author. Mailing address: Department of Neuroscience (D13), Osaka University Graduate School of Medicine, Yamadaoka 2-2, Suita, Osaka 565-0871, Japan. Phone: 81 6 6879 3680. Fax: 81 6 6879 3689. E-mail: sobue{at}nbiochem.med.osaka-u.ac.jp.

{triangledown} Published ahead of print on 9 October 2006.

{dagger} Present address: Department of Laboratory Medicine, Ehime University School of Medicine, Toon-shi, Ehime 791-0295, Japan.

Molecular and Cellular Biology, December 2006, p. 9456-9470, Vol. 26, No. 24
0270-7306/06/$08.00+0     doi:10.1128/MCB.00759-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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