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J. Biol. Chem., Vol. 277, Issue 32, 28683-28689, August 9, 2002

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The Phosphorylation Site Located in the A Region of Retinoic X Receptor  Is Required for the Antiproliferative Effect of Retinoic Acid (RA) and the Activation of RA Target Genes in F9 Cells^*

Julie Bastien, Sylvie Adam-Stitah, Jean-Luc Plassat, Pierre Chambon, and Cécile Rochette-Egly^§

From the Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP/Collège de France, BP 163, 67404 Illkirch Cedex, France

Mouse F9 embryocarcinoma cells constitute a well established cell autonomous model system for investigating retinoic acid (RA) signaling in vitro. RA induces the differentiation of F9 cells grown as monolayers into endodermal-like cells and decreases their rate of proliferation. Knock-out of the retinoic X receptor  (RXR) gene abolishes endodermal differentiation and the induction of several endogenous RA-responsive genes. RXR null cells are also drastically impaired in their antiproliferative response to RA. The role of the RXR phosphorylation site located in the N-terminal A region (Ser²²) has been investigated here by establishing cell lines re-expressing RXR either wild type or mutated at the phosphorylation site (RXRS22A) in a RXR-null background. We show that Ser²² is dispensable for RA-induced endodermal differentiation but is crucial for the expression of several RA-responsive genes. Ser²² is also indispensable for the antiproliferative effect of RA and necessary for the RA-induced down-regulation of p21^CIP and p27^KIP CKIs proteins that are known to be involved in the control of cell cycle progression.

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