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J. Biol. Chem., Vol. 280, Issue 19, 18710-18716, May 13, 2005

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Mint Represses Transactivation of the Type II Collagen Gene Enhancer through Interaction with A-crystallin-binding Protein 1^*

Xi Yang, Junfeng Li, Hongyan Qin, Hui Yang, Junlin Li, Peng Zhou, Yingmin Liang, and Hua Han

From the Department of Medical Genetics and Developmental Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xian 710032, China

Collagen type II is an extracellular matrix protein important for cartilage and bone formation, and its expression is controlled by multiple cis- and trans-acting elements, including the zinc finger transcription factor A-crystallin-binding protein 1 (CRYBP1). Here we show that MSX2-interacting nuclear target protein (MINT), a conserved transcriptional repressor, associates with CRYBP1 and negatively regulates the transactivation of the collagen type II gene (Col2a1) enhancer. We identified CRYBP1 as a binding partner of MINT by screening a mouse embryonic cDNA library using the yeast two-hybrid system. We demonstrated that the C terminus of MINT interacts with the C terminus of CRYBP1 using the mammalian cell two-hybrid assay, glutathione S-transferase pull-down, and co-immunoprecipitation analyses. Furthermore, MINT and CRYBP1 form a complex on the Col2a1 enhancer, as shown by chromatin immunoprecipitation and gel shift assays. In the presence of CRYBP1, overexpression of MINT or its C-terminal fragment in cells repressed a reporter construct driven by the Col2a1 enhancer elements. This transcription repression is dependent on histone deacetylase, the main co-repressor recruited by MINT. The present study shows that MINT is involved in CRYBP1-mediated Col2a1 gene repression and may play a role in regulation of cartilage development.

Received for publication, January 24, 2005 , and in revised form, March 11, 2005.

^* This work was supported by the National Natural Science Foundation of China (Grants 30300110, 30330550, and 30425015) and the Ministry of Science and Technology of China (Grant 001CB509906), and the Program for Changjiang Scholars and Innovative Research Team in the University of the Ministry of Education of China. 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.

To whom correspondence should be addressed: Dept. of Medical Genetics and Developmental Biology, Fourth Military Medical University, Changle Xi St. #17, Xian 710032, China. Tel.: 86-29-8337-4487; Fax: 86-29-8324-6270; E-mail: huahan{at}fmmu.edu.cn.

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