Aryl hydrocarbon receptor (AhR) regulates adipocyte differentiation by assembling CRL4B ubiquitin ligase to target PPARγ for proteasomal degradation

• Published in: The Journal of biological chemistry 2019-11, Vol.294 (48), p.18504-18515
• Main Authors: Dou, Hao, Duan, Yuyao, Zhang, Xiaohui, Yu, Qian, Di, Qian, Song, Yu, Li, Peishan, Gong, Yaoqin
• Format: Article
• Language: English
• Subjects: 3T3-L1 Cells; adipocyte differentiation; Adipocytes - cytology; Adipocytes - metabolism; Animals; aryl hydrocarbon receptor (AhR) (AHR); Cell Differentiation; cullin 4B; Cullin Proteins - genetics; Cullin Proteins - metabolism; E3 ubiquitin ligase; HEK293 Cells; Humans; Male; Mice; Mice, Knockout; peroxisome proliferator-activated receptor (PPAR); peroxisome proliferator-activated receptor γ (PPARγ); PPAR gamma - genetics; PPAR gamma - metabolism; Proteasome Endopeptidase Complex - metabolism; protein degradation; Protein Synthesis and Degradation; Receptors, Aryl Hydrocarbon - genetics; Receptors, Aryl Hydrocarbon - metabolism; RNA Interference; Ubiquitination; ubiquitylation (ubiquitination)
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA119.009282

Peroxisome proliferator-activated receptor γ (PPARγ) is the central regulator of adipogenesis, and its dysregulation is linked to obesity and metabolic diseases. Identification of the factors that regulate PPARγ expression and activity is therefore crucial for combating obesity. Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor with a known role in xenobiotic detoxification. Recent studies have suggested that AhR also plays essential roles in energy metabolism. However, the detailed mechanisms remain unclear. We previously reported that experiments with adipocyte-specific Cullin 4b (Cul4b)-knockout mice showed that CUL4B suppresses adipogenesis by targeting PPARγ. Here, using immunoprecipitation, ubiquitination, real-time PCR, and GST-pulldown assays, we report that AhR functions as the substrate receptor in CUL4B-RING E3 ubiquitin ligase (CRL4B) complex and is required for recruiting PPARγ. AhR overexpression reduced PPARγ stability and suppressed adipocyte differentiation, and AhR knockdown stimulated adipocyte differentiation in 3T3-L1 cells. Furthermore, we found that two lysine sites on residues 268 and 293 in PPARγ are targeted for CRL4B-mediated ubiquitination, indicating cross-talk between acetylation and ubiquitination. Our findings establish a critical role of AhR in regulating PPARγ stability and suggest that the AhR–PPARγ interaction may represent a potential therapeutic target for managing metabolic diseases arising from PPARγ dysfunction.