The forkhead transcription factor FOXO1 stimulates the expression of the adipocyte resistin gene

• FOXO1 and resistin mRNA levels are higher in differentiating preadipocytes than those in non-differentiating preadipocytes.
• FOXO1 activates resistin promoter activity in preadipocytes and adipocytes.
• FOXO1 activation of resistin promoter depends on the phosphorylation and dephosphorylation of FOXO1.

Resistin is known as an adipocyte-specific hormone that can cause insulin resistance and decrease adipocyte differentiation. It can be regulated by transcriptional factors, but the possible role of forkhead transcription factor FOXO1 in regulating resistin gene expression is still unknown. Using 3T3 fibroblast and C3H10T1/2 and 3T3-L1 adipocytes, we found that transient overexpression of a non-phosphorylatable, constitutively active FOXO1, but not the wild type of FOXO1 or a DNA binding-deficient FOXO1, activated resistin promoter-directed luciferase expression. However, transient overexpression of a dominant-negative FOXO1 inactivated resistin promoter activity and reduced resistin mRNA expression. These observations indicate that the action of FOXO1 on resistin gene expression requires the activation of FOXO1 and that the effect of FOXO1 depends on the phosphorylation and dephosphorylation of FOXO1. The FOXO1 protein target sites on the resistin promoter were localized to the proximal −3545 to −787 bp of 5′-flanking region of the resistin promoter. A chromatin immunoprecipitation assay also showed that FOXO1 bound the resistin promoter at nucleotide regions of −1539 to −1366 bp and −1016 to −835 bp, but not at the regions of −795 to −632 bp. Results of this study suggest that FOXO1 transcription factor likely activates the expression of adipocyte resistin gene via direct association with the upstream resistin promoter.