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J. Biol. Chem., Vol. 281, Issue 45, 33997-34008, November 10, 2006

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Identification and Characterization of a Novel Gene, Mcpr1, and Its Possible Function in the Proliferation of Embryonic Palatal Mesenchymal Cells^*

Dong-Ying Xuan, Xin Li, Zhi-Hong Deng^¶, Hua-Li Zhang, Pei-xun Feng, Xiao-Yan Duan, and Yan Jin¹

From the Department of Oral Histology and Pathology, College of Stomatology, the Research and Development Center for Tissue Engineering, and the ^¶Department of Otolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, China

We cloned a novel mouse cDNA, Mcpr1 (mouse cleft palate-related gene 1), between retinoic acid (RA)-treated murine embryonic palatal and control shelves by improved subtractive hybridization. Its transcript was identified by Northern blotting. The open reading frame encodes 132 amino acids and shows almost no identity to other genetic products. Mcpr1 expression could be detected extensively in adult mouse tissues and during murine embryonic development. It was identified to be significantly stimulated by RA in murine palatal shelves at embryonic day 12 and in palatal mesenchymal cells in vitro. We demonstrate that MCPR1 protein was localized primarily in the cytoplasm and could be synthesized and secreted by transfected COS-7 cells. Both the secretory and recombinant proteins of Mcpr1 inhibited proliferation of murine embryonic palatal mesenchymal cells and impeded the progression from the G₁ to S phase in the cell cycle. The cells were prone to apoptosis after exposure to glutathione S-transferase-MCPR1. Furthermore, knockdown of MCPR1 protein levels by antisense oligodeoxynucleotides promoted progression of cells from the G₁ to S phase and completely abolished the RA-induced block of the cell cycle from the G₁ to S phase. These findings suggest that Mcpr1 might function as one of the RA-up-regulated genes involved in inhibiting cell proliferation during palatogenesis and RA-induced cleft palate by regulating proliferation and apoptosis of embryonic palatal mesenchymal cells and might even play a role in the development of many other organs.

Received for publication, June 7, 2006 , and in revised form, September 8, 2006.

^* This work was supported by Grants 2002AA205041 and 2005AA205241 from Development of High and New Science and Technology Project 863 and by Project 30572046 of the Nature Science Foundation 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.

The nucleotide sequence(s) reported in this paper has been submitted to the Gen-Bank^TM/EBI Data Bank with accession number(s) AY074887 [GenBank] .

¹ To whom correspondence should be addressed: Dept. of Oral Histology and Pathology, College of Stomatology, Fourth Military Medical University, 7 Kangfu Rd., Xi'an, Shaanxi 710032, China. Tel.: 86-29-8477-6147; Fax: 86-29-8321-8039; E-mail: yanjin{at}fmmu.edu.cn or yanjinfmmu{at}vip.sina.com.

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