LIM protein Ajuba functions as a nuclear receptor corepressor and negatively regulates retinoic acid signaling

• Published in: Proceedings of the National Academy of Sciences - PNAS 2010-02, Vol.107 (7), p.2938-2943
• Main Authors: Hou, Zhaoyuan, Peng, Hongzhuang, White, David E, Negorev, Dmitri G, Maul, Gerd G, Feng, Yunfeng, Longmore, Gregory D, Waxman, Samuel, Zelent, Arthur, Rauscher, Frank J. III
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs - genetics; Amino Acid Motifs - physiology; Animals; Binding sites; Biological Sciences; Blotting, Western; Cell culture techniques; Cell Line; Cell lines; Cellular differentiation; Cellular immunity; Chromatin; Chromatin Immunoprecipitation; Co repressor proteins; Co-Repressor Proteins - metabolism; Embryonic stem cells; Gene Expression Regulation - physiology; HEK293 cells; Homeodomain Proteins - metabolism; Humans; Immunoprecipitation; LIM Domain Proteins; Luciferases; Mice; Microscopy, Fluorescence; Molecules; Nuclear interactions; Protein Binding; Protein-Arginine N-Methyltransferases - metabolism; Proteins; Receptors; Receptors, Cytoplasmic and Nuclear - metabolism; Receptors, Retinoic Acid - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Signal transduction; Signal Transduction - physiology; Stem cells; T cell receptors; Tretinoin - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0908656107

Corepressors play an essential role in nuclear receptor-mediated transcriptional repression. In general, corepressors directly bind to nuclear receptors via CoRNR boxes (L/I-X-X-I/V-I) in the absence of ligand and appear to act as scaffolds to further recruit chromatin remodeling complexes to specific target genes. Here, we describe the identification of the multiple LIM domain protein Ajuba as a unique corepressor for a subset of nuclear hormone receptors. Ajuba contains functional nuclear-receptor interacting motifs and selectively interacts with retinoic acid receptors (RARs) and rexinoid receptor (RXRs) subtypes in a ligand-dependent manner. Simultaneous mutation of these motifs abolishes RAR binding and concomitantly leads to loss of repression on RARE reporter genes. P19 cells depleted of Ajuba are highly sensitized to all-trans retinoic acid (atRA)-induced transcription and differentiation. In the absence of atRA, Ajuba can be readily found at the RARE control elements of RAR endogenous target genes. Stimulation of cells with atRA results in the dissociation of Ajuba from these regions. Moreover, we observed that coexpression of the known Ajuba binding partner Prmt5 (protein arginine methyltransferase-5) inhibited the Ajuba/RAR interaction. The high-affinity Ajuba-RAR/RXR interaction site overlaps the region responsible for Ajuba/Prmt5 binding, and thus binding appears to be mutually exclusive, providing a potential mechanism for these observations. Identification of Ajuba as a unique corepressor for nuclear receptors sheds new light on mechanisms for nuclear receptor-mediated repression and provides a unique target for developing more effective therapeutics to modulate this important pathway.