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Reactive Oxygen Species Facilitate Adipocyte Differentiation by Accelerating Mitotic Clonal Expansion*

By Haemi Lee, Yoo Jeong Lee, Hyeonjin Choi, Eun Hee Ko and Jae-woo Kim


Growth-arrested 3T3-L1 preadipocytes rapidly express CCAAT/enhancer-binding protein-β (C/EBPβ) upon hormonal induction of differentiation. However, the DNA binding activity of C/EBPβ is not activated until the cells synchronously reenter S phase during the mitotic clonal expansion (MCE) phase of differentiation. In this period, C/EBPβ is sequentially phosphorylated by MAPK and glycogen synthase kinase-3β, inducing C/EBPβ DNA binding activity and transcription of its target genes. Because the DNA binding activity of C/EBPβ is further enhanced by oxidation in vitro, we investigated how redox state affects C/EBPβ DNA binding and MCE during adipogenesis. When 3T3-L1 cells were treated with H2O2 and hormonal stimuli, differentiation was accelerated with increased expression of peroxisome proliferator-activated receptor γ. Interestingly, cell cycle progression (S to G2/M phase) was markedly enhanced by H2O2, whereas antioxidants caused an S phase arrest during the MCE. H2O2 treatment resulted in the early appearance of a punctate pattern observed by immunofluorescent staining of C/EBPβ, which is a hallmark for C/EBPβ binding to regulatory elements, whereas a short antioxidant treatment rapidly dispersed the centromeric localization of C/EBPβ. Consistently, reactive oxygen species production was increased during 3T3-L1 differentiation. Our results indicate that redox-induced C/EBPβ DNA binding activity, along with the dual phosphorylation of C/EBPβ, is required for the MCE and terminal differentiation of adipocytes

Topics: Metabolism and Bioenergetics
Publisher: American Society for Biochemistry and Molecular Biology
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Provided by: PubMed Central
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