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Originally published In Press as doi:10.1074/jbc.M203157200 on May 22, 2002

J. Biol. Chem., Vol. 277, Issue 33, 29840-29846, August 16, 2002
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Negative Regulation of Bone Morphogenetic Protein/Smad Signaling by Cas-interacting Zinc Finger Protein in Osteoblasts*

Zhong-Jian ShenDagger , Tetsuya Nakamoto§, Kunikazu TsujiDagger , Akira NifujiDagger , Kohei Miyazono, Toshihisa Komori||, Hisamaru Hirai§, and Masaki NodaDagger **

From the Dagger  Department of Molecular Pharmacology, Medical Research Institute, Tokyo Medical and Dental University, 3-10 Kanda-Sunugadai 2-Chome Chiyoda-ku, Tokyo, 101 Japan, the § Department of Hematology and Oncology, and  Department of Molecular Pathology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan, and the || Department of Molecular Medicine, Osaka University Medical School, Osaka, Japan

Bone morphogenetic protein (BMP) signaling regulates body axis determination, apoptosis, and differentiation of various types of cells including neuron, gut, and bone cells. However, the molecules involved in such BMP regulation of biological events have not been fully understood. Here, we examined the involvement of Cas-interacting zinc finger protein (CIZ) in the modulation of BMP2-induced osteoblastic cell differentiation. CIZ overexpression in osteoblastic MC3T3E1 cells suppressed BMP2-enhanced expression of alkaline phosphatase, osteocalcin, and type I collagen genes. Upstream analyses revealed that CIZ overexpression also suppressed BMP2-induced enhancement of the mRNA expression of Cbfa1, which is a critical transcription factor for osteoblastic differentiation. BMP-induced Smad1 and Smad5 activation of GCCG-mediated transcription was blocked in the presence of CIZ overexpression. CIZ overexpression alone in the absence of BMP2 moderately enhanced basal levels of Cbfa1 mRNA expression. CIZ overexpression also enhanced 1.8-kb Cbfa1 promoter activity in the absence of BMP2, whereas it suppressed the promoter activity in the presence of BMP2. Finally, CIZ overexpression suppressed the formation of mineralized nodules in osteoblastic cell cultures. These data indicate that CIZ is a novel type inhibitor of BMP/Smad signaling.

* This research was supported by the grants-in-aid received from the Japanese Ministry of Education (14207056, 14034214, 14028022, 12557123, 13045011, and 13216034), grants from NASDA, Japan Society for Promotion of Science (Research for the Future Program, Genome Science), and Tokyo Biochemistry Research Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The 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.: 81-3-528-8066; Fax: 81-3-5280-8066; E-mail: noda.mph@mri.tmd.ac.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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