Induction of DEAD Box helicase 5 in early adipogenesis is regulated by Ten-eleven translocation 2

Dead box helicase 5 (DDX5) is an RNA helicase that is has cellular function on RNA splicing and transcriptional regulation. It has been reported to be involved in cell differentiation including adipogenesis. However, it is not clear how DDX5 is regulated during adipogenesis. Our previous report demonstrated that the Ten-eleven translocation methyl-cytosine dioxygenase 2 (TET2) is required for adipogenesis. This study was aimed to investigate DDX5 as a direct target of TET2 upon adipogenic induction of 3T3-L1 preadipocyte. Microarray-based screening of differentially expressed genes upon TET2 knockdown identified genes involved in cell cycle, DNA replication, and ribosome biology as major targets of TET2 in the initial step of adipogenic induction. The Ddx5 gene was identified and validated as the target. TET2-mediated epigenetic regulation of the Ddx5 gene was measured by two independent methods including immunoprecipitation against 5-hydroxymethylcytosine (5hmC) and 5-methylcytosine (5mC) as well as EpiMark 5hmC and 5mC analysis. Ddx5 expression was downregulated upon TET2 knockdown, coincided with a significant decrease of 5hmC at the Ddx5 locus. DDX5 knockdown significantly suppressed adipogenesis, while DDX5 overexpression promoted it. Importantly, DDX5 overexpression, when co-transfected, rescued the process of adipogenesis, which was hindered by TET2 siRNA treatment. The findings suggest TET2-mediated regulation of the Ddx5 gene is required for an initial step of adipogenesis.