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J. Biol. Chem., Vol. 280, Issue 5, 3166-3177, February 4, 2005

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Regulation of Chicken ccn2 Gene by Interaction between RNA cis-Element and Putative trans-Factor during Differentiation of Chondrocytes^*

Yoshiki Mukudai, Satoshi Kubota, Takanori Eguchi¶, Seiji Kondo¶, Kyouji Nakao, and Masaharu Takigawa||

From the Biodental Research Center, Okayama University Dental School and Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8525 and the ^¶Postdoctoral Fellowships of Japanease Society for the Promotion of Science, 6 Ichibancho, Chiyoda-ku, Tokyo, 102-8471, Japan

CCN2/CTGF is a multifunctional growth factor. Our previous studies have revealed that CCN2 plays important roles in both growth and differentiation of chondrocytes and that the 3'-untranslated region (3'-UTR) of ccn2 mRNA contains a cis-repressive element of gene expression. In the present study, we found that the stability of chicken ccn2 mRNA is regulated in a differentiation stage-dependent manner in chondrocytes. We also found that stimulation by bone morphogenetic protein 2, platelet-derived growth factor, and CCN2 stabilized ccn2 mRNA in proliferating chondrocytes but that it destabilized the mRNA in prehypertrophic-hypertrophic chondrocytes. The results of a reporter gene assay revealed that the minimal repressive cis-element of the 3'-UTR of chicken ccn2 mRNA was located within the area between 100 and 150 bases from the polyadenylation tail. Moreover, the stability of ccn2 mRNA was correlated with the interaction between this cis-element and a putative 40-kDa trans-factor in nuclei and cytoplasm. In fact, the binding between them was prominent in proliferating chondrocytes and attenuated in (pre)hypertrophic chondrocytes. Stimulation by the growth factors repressed the binding in proliferating chondrocytes; however, it enhanced it in (pre)hypertrophic chondrocytes. Therefore, gene expression of ccn2 mRNA during endochondral ossification is properly regulated, at least in part, by changing the stability of the mRNA, which arises from the interaction between the RNA cis-element and putative trans-factor.

Received for publication, October 13, 2004 , and in revised form, November 16, 2004.

^* This work was supported by grants from the programs Grants-inaid for Scientific Research (S) (to M. T.) and (C) (to S. Kubota) and Grants-in-aid for Exploratory Research (to M. T.) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and by grants from the Sumitomo Foundation (to M. T.) and the Nakatomi Foundation (to S. Kubota). 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.

This report is dedicated to the memory of Dr. Hitoshi Akedo.

^|| To whom correspondence should be addressed. Tel.: 81-86-235-6645; Fax: 81-86-235-6649; E-mail: takigawa{at}md.okayama-u.ac.jp.

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				Y. Mukudai, S. Kubota, H. Kawaki, S. Kondo, T. Eguchi, K. Sumiyoshi, T. Ohgawara, T. Shimo, and M. Takigawa Posttranscriptional Regulation of Chicken ccn2 Gene Expression by Nucleophosmin/B23 during Chondrocyte Differentiation Mol. Cell. Biol., October 1, 2008; 28(19): 6134 - 6147. [Abstract] [Full Text] [PDF]

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