Despouy Gilles, Bastie Jean-Nöel, Deshaies Sylvie, Balitrand Nicole, Mazharian Alexandra, Rochette-Egly Cécile, Chomienne Christine, Delva Laurent
Laboratoire de Biologie Cellulaire Hématopoiétique, Equipe Mixte Inserm 00-03, Institut Universitaire d'Hématologie, Hôpital Saint-Louis, 1 Avenue Claude Vellefaux, Paris 75010, USA.
J Biol Chem. 2003 Feb 21;278(8):6355-62. doi: 10.1074/jbc.M210697200. Epub 2002 Dec 12.
Ligand-induced transcription activation of retinoic acid (RA) target genes by nuclear receptors (retinoic acid (RAR) and retinoid X (RXR) receptors) depends on the recruitment of coactivators. We have previously demonstrated that the small 15-kDa cellular RA-binding protein II (CRABPII) is a coactivator present in the RA-dependent nuclear complex. As identifying cell-specific partners of CRABPII might help to understand the novel control of RA signaling, we performed a yeast two-hybrid screen of a hematopoietic HL-60 cDNA library using human CRABPII as bait and have subsequently identified human cyclin D3 as a partner of CRABPII. Cyclin D3 interacted with CRABPII in a ligand-independent manner and equally bound RAR alpha, but not RXR alpha, and only in the presence of RA. We further show that cyclin D3 positively modulated RA-mediated transcription through CRABPII. Therefore, cyclin D3 may be part of a ternary complex with CRABPII and RAR. Finally, we show that cyclin D3 expression paralleled HL-60 differentiation and arrest of cell growth. These findings led us to speculate that control of cell proliferation during induction of differentiation may directly involve, at the transcriptional level, nuclear receptors, coactivators, and proteins of the cell cycle in a cell- and nuclear receptor-specific manner.
核受体(视黄酸受体(RAR)和类视黄醇X受体(RXR))通过配体诱导的视黄酸(RA)靶基因转录激活依赖于共激活因子的募集。我们之前已经证明,小的15 kDa细胞视黄酸结合蛋白II(CRABPII)是存在于RA依赖性核复合物中的一种共激活因子。由于鉴定CRABPII的细胞特异性伙伴可能有助于理解RA信号传导的新调控机制,我们以人CRABPII为诱饵对造血HL-60 cDNA文库进行了酵母双杂交筛选,随后鉴定出人细胞周期蛋白D3是CRABPII的伙伴。细胞周期蛋白D3以不依赖配体的方式与CRABPII相互作用,并且同样与RARα结合,但不与RXRα结合,且仅在有RA存在时结合。我们进一步表明,细胞周期蛋白D3通过CRABPII正向调节RA介导的转录。因此,细胞周期蛋白D3可能是与CRABPII和RAR形成三元复合物的一部分。最后,我们表明细胞周期蛋白D3的表达与HL-60细胞的分化和细胞生长停滞平行。这些发现使我们推测,在分化诱导过程中对细胞增殖的控制可能在转录水平上以细胞和核受体特异性的方式直接涉及核受体、共激活因子和细胞周期蛋白。