miR-199a*, a Bone Morphogenic Protein 2-responsive MicroRNA, Regulates Chondrogenesis via Direct Targeting to Smad1

被引:203
作者
Lin, Edward A. [1 ]
Kong, Li [1 ]
Bai, Xiao-Hui [1 ]
Luan, Yi [1 ]
Liu, Chuan-ju [1 ,2 ]
机构
[1] NYU, Sch Med, Dept Orthopaed Surg, New York, NY 10003 USA
[2] NYU, Sch Med, Dept Cell Biol, New York, NY 10016 USA
基金
新加坡国家研究基金会; 美国国家卫生研究院;
关键词
OLIGOMERIC MATRIX PROTEIN; INDUCIBLE P204 PROTEIN; OSTEOGENIC DIFFERENTIATION; TRANSCRIPTION FACTOR; GENE-EXPRESSION; OSTEOBLAST DIFFERENTIATION; MICROMASS CULTURES; MESSENGER-RNA; STEM-CELLS; SKELETOGENESIS;
D O I
10.1074/jbc.M807709200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
MicroRNAs (miRNA) are short non-coding RNA molecules that regulate a variety of biological processes. The role of miRNAs in BMP2-mediated biological processes is of considerable interest. A comparative miRNA array led to the isolation of several BMP2-responsive miRNAs. Among them, miR-199a* is of particular interest, because it was reported to be specifically expressed in the skeletal system. Here we demonstrate that miR-199a* is an early responsive target of BMP2: its level was dramatically reduced at 5 h, quickly increased at 24 h and remained higher thereafter in the course of BMP2-triggered chondrogenesis of a micromass culture of pluripotent C3H10T1/2 stem cells. miR-199a* significantly inhibited early chondrogenesis, as revealed by the reduced expression of early marker genes for chondrogenesis such as cartilage oligomeric matrix protein (COMP), type II collagen, and Sox9, whereas anti-miR-199a* increased the expression of these chondrogenic marker genes. A computer-based prediction algorithm led to the identification of Smad1, a well established downstream molecule of BMP-2 signaling, as a putative target of miR-199a*. The pattern of Smad1 mRNA expression exhibited the mirror opposite of miR-199a* expression following BMP-2 induction. Furthermore, miR-199a* demonstrated remarkable inhibition of both endogenous Smad1 as well as a reporter construct bearing the 3-un-translated region of Smad1 mRNA. In addition, mutation of miR-199a* binding sites in the 3'-untranslated region of Smad1 mRNA abolished miR-199a*-mediated repression of reporter gene activity. Mechanism studies revealed that miR-199a* inhibits Smad1/Smad4-mediated transactivation of target genes, and that overexpression of Smad1 completely corrects miR-199a*-mediated repression of early chondrogenesis. Taken together, miR-199a* is the first BMP2 responsive microRNA found to adversely regulate early chondrocyte differentiation via direct targeting of the Smad1 transcription factor.
引用
收藏
页码:11326 / 11335
页数:10
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