Regulation of Adipocyte Differentiation by METTL4, a 6 mA Methylase

As one of the most abundant DNA methylation form in prokaryotes, N⁶-methyladenine nucleotide (6 mA) was however only recently identified in eukaryotic genomes. To explore the implications of N⁶-adenine methylation in adipogenesis, genomic N⁶-adenine methylation was examined across adipocyte differentiation stages of 3T3-L1 cells. When the N⁶-adenine methylation profiles were analyzed and compared with the levels of gene expression, a positive correlation between the density of promoter 6 mA and gene expression level was uncovered. By means of in vitro methylation and gene knockdown assay, METTL4, a homologue of Drosophila methylase CG14906 and C. elegans methylase DAMT-1, was demonstrated to be a mammalian N⁶-adenine methylase that functions in adipogenesis. Knockdown of Mettl4 led to altered adipocyte differentiation, shown by defective gene regulation and impaired lipid production. We also found that the effects of N⁶-adenine methylation on lipid production involved the regulation of INSR signaling pathway, which promotes glucose up-taking and lipid production in the terminal differentiation stage.