PBX proteins are important mediators for retinoic acid-dependent endodermal and neuronal differentiation of mouse embryonal carcinoma P19 cells

doi:10.1074/jbc.M313938200

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Papers In Press, published online ahead of print January 23, 2004
J. Biol. Chem, 10.1074/jbc.M313938200
Submitted on December 19, 2003
Revised on January 14, 2004
Accepted on January 23, 2004

PBX proteins are important mediators for retinoic acid-dependent endodermal and neuronal differentiation of mouse embryonal carcinoma P19 cells

Pu Qin, Juliet M. Haberbusch, Zhenping Zhang, Kenneth J. Soprano, and Dianne Robert Soprano

Biochemistry Dept., Temple University School of Medicine, Philadelphia, PA 19140

Corresponding Author: dsoprano{at}temple.edu

Pre-B cell leukemia transcription factors (PBXs) act as co-factors in the transcriptional regulation mediated by HOX proteins during embryonic development and cellular differentiation. PBX1 protein is expressed throughout murine embryonic development and its deletion in mice disrupts chondrogenesis. PBX protein levels are also increased in mouse embryonal carcinoma P19 cells during retinoic acid (RA)-induced differentiation. To elucidate the role of PBX proteins in this process, we stably overexpressed PBX1b antisense mRNA in P19 cells (PBX1b-AS cells). PBX1b-AS cells did not differentiate to neuronal or endodermal cells following treatment with RA suggesting PBX proteins are required for both processes. Furthermore, we demonstrated that PBX proteins regulate the RA-dependent induction in the mRNA levels of bone morphogenesis protein 4 (BMP4) and Decorin (DCN) in P19 cells using both PBX1b-AS cells and PBX1 siRNA. Chromatin immunoprecipitation assays further demonstrate that PBX proteins directly bind to the promoter of BMP4 and DCN genes in vivo in a RA-dependent fashion. In addition, type I and type II BMP receptor mRNAs levels were also increased in P19 cells following RA treatment however this was PBX-independent. Taken together these data demonstrate that PBX proteins are required for RA-induced differentiation of P19 cells and that PBX proteins regulate the expression of BMP4 and DCN during this differentiation process.

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