title: Ubiquitination of Lysine 867 of the Human SETDB1 Protein Upregulates Its Histone H3 Lysine 9 (H3K9) Methyltransferase Activity.
abstract: Posttranslational modifications (PTMs) of proteins play a crucial role in regulating protein-protein interactions, enzyme activity, subcellular localization, and stability of the protein. SET domain, bifurcated 1 (SETDB1) is a histone methyltransferase that regulates the methylation of histone H3 on lysine 9 (H3K9), gene silencing, and transcriptional repression. The C-terminal region of SETDB1 is a key site for PTMs, and is essential for its enzyme activity in mammalian and insect cells. In this study, we aimed to evaluate more precisely the effect of PTMs on the H3K9 methyltransferase activity of SETDB1. Using mass spectrometry analysis, we show that the C-terminal region of human SETDB1 purified from insect cells is ubiquitinated. We also demonstrate that the ubiquitination of lysine 867 of the human SETDB1 is necessary for full H3K9 methyltransferase activity in mammalian cells. Finally, we show that SETDB1 ubiquitination regulates the expression of its target gene, serpin peptidase inhibitor, clade E, member 1 (SERPINE1) by methylating H3K9. These results suggest that the ubiquitination of SETDB1 at lysine 867 controls the expression of its target gene by activating its H3K9 methyltransferase activity.
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.|Laboratory for System Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Kenji
authors: K
authors: Ishimoto
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Natsuko
authors: N
authors: Kawamata
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Yoshie
authors: Y
authors: Uchihara
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Moeka
authors: M
authors: Okubo
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Reiko
authors: R
authors: Fujimoto
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Eiko
authors: E
authors: Gotoh
authors: Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
authors: Keisuke
authors: K
authors: Kakinouchi
authors: Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
authors: Eiichi
authors: E
authors: Mizohata
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Nobumasa
authors: N
authors: Hino
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Yoshiaki
authors: Y
authors: Okada
authors: Department of Quantitative Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Yasuhiro
authors: Y
authors: Mochizuki
authors: Laboratory for System Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Toshiya
authors: T
authors: Tanaka
authors: Department of Quantitative Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Takao
authors: T
authors: Hamakubo
authors: Division of Metabolic Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Juro
authors: J
authors: Sakai
authors: Laboratory for System Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Tokyo, Japan.
authors: Tatsuhiko
authors: T
authors: Kodama
authors: Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
authors: Tsuyoshi
authors: T
authors: Inoue
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Keisuke
authors: K
authors: Tachibana
authors: Laboratory of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors: Takefumi
authors: T
authors: Doi
mesh_terms: D000818:Animals; D002460:Cell Line; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008954:Models, Biological; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011496:Protein Methyltransferases / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D006657:Histones; D000076983:Histone Methyltransferases; D011496:Protein Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C454062:SETDB1 protein, human; D008239:Lysine