Roles of 3'UTR and IDR in regulating activity and chromatin localization of JMJD3 [ChIP-seq]

JMJD3 cellular activity can be regulated by altered chromatin localization to genomic targets or by protein conformation changes that alter its catalytic activity. We found that the 3'UTR in the KDM6B mRNA template is required for the JMJD3 protein to achive full demethylase activity in cells and that 3'UTR dependent regulation requires its intrinsically disordered region (IDR). In order to dissect the mechanism underlying 3'UTR- and IDR-coordinated control of JMJD3 protein activity in cells, we used ChIP-seq to identify localization and demethylase activity of full-length JMJD3 expressed from constructs with or without its 3'UTR and a IDR-deleted mutant. ChIP-seq of HA-tagged KDM6B/JMJD3 and H3K27me3 in HeLa cells transfected with HA-tagged cDNA vectors expressing full-length KDM6B coding sequence (-NU), full-length KDM6B coding sequence and its 3′UTR, or IDR-deleted KDM6B coding sequence (IDRdel-NU). Transfected cells were enriched with FACS.