| First Author | Watanabe Y | Year | 2020 |
| Journal | FASEB J | Volume | 34 |
| Issue | 4 | Pages | 5827-5837 |
| PubMed ID | 32141127 | Mgi Jnum | J:305214 |
| Mgi Id | MGI:6695426 | Doi | 10.1096/fj.201902575R |
| Citation | Watanabe Y, et al. (2020) Protein S-glutathionylation stimulate adipogenesis by stabilizing C/EBPbeta in 3T3L1 cells. FASEB J 34(4):5827-5837 |
| abstractText | Reactive oxygen species (ROS) increase during adipogenesis and in obesity. Oxidants react with cysteine residues of proteins to form glutathione (GSH) adducts, S-glutathionylation, that are selectively removed by glutaredoxin-1 (Glrx). We have previously reported that Glrx knockout mice had increased protein S-glutathionylation and developed obesity by an unknown mechanism. In this study, we demonstrated that 3T3L1 adipocytes differentiation increased ROS and protein S-glutathionylation. Glrx ablation elevated protein S-glutathionylation and lipid content in 3T3L1 cells. Glrx replenishment decreased the lipid content of Glrx KO 3T3L1 cells. Glrx KO also increased protein expression and protein S-glutathionylation of the adipogenic transcription factor CCAAT enhancer-binding protein (C/EBP) beta. Protein S-glutathionylation decreased the interaction of C/EBPbeta and protein inhibitor of activated STAT (PIAS) 1, a small ubiquitin-related modifier E3 ligase that facilitates C/EBPbeta degradation. Experiments with truncated mutant C/EBPbeta demonstrated that PIAS1 interacted with the liver-enriched inhibitory protein (LIP) region of C/EBPbeta. Furthermore, mass spectrometry analysis identified protein S-glutathionylation of Cys201 and Cys296 in the LIP region of C/EBPbeta. The C201S, C296S double-mutant C/EBPbeta prevented protein S-glutathionylation and preserved the interaction with PIAS1. In summary, Glrx ablation stimulated 3T3L1 cell differentiation and adipogenesis via increased protein S-glutathionylation of C/EBPbeta, stabilizing and increasing C/EBPbeta protein levels. |
