Journal
FASEB JOURNAL
Volume 34, Issue 4, Pages 5827-5837Publisher
WILEY
DOI: 10.1096/fj.201902575R
Keywords
CCAAT enhancer-binding protein; glutaredoxin-1; reactive oxygen species; S-glutathionylation
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Funding
- NHLBI NIH HHS [R01 HL133013] Funding Source: Medline
- NIDDK NIH HHS [R01 DK103750] Funding Source: Medline
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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/EBP beta and protein inhibitor of activated STAT (PIAS) 1, a small ubiquitin-related modifier E3 ligase that facilitates C/EBP beta degradation. Experiments with truncated mutant C/EBP beta demonstrated that PIAS1 interacted with the liver-enriched inhibitory protein (LIP) region of C/EBP beta. Furthermore, mass spectrometry analysis identified protein S-glutathionylation of Cys201 and Cys296 in the LIP region of C/EBP beta. The C201S, C296S double-mutant C/EBP beta 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/EBP beta, stabilizing and increasing C/EBP beta protein levels.
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