TY - JOUR
T1 - Ubiquitination of lysine 867 of the human SETDB1 protein upregulates its histone H3 lysine 9 (H3K9) methyltransferase activity
AU - Ishimoto, Kenji
AU - Kawamata, Natsuko
AU - Uchihara, Yoshie
AU - Okubo, Moeka
AU - Fujimoto, Reiko
AU - Gotoh, Eiko
AU - Kakinouchi, Keisuke
AU - Mizohata, Eiichi
AU - Hino, Nobumasa
AU - Okada, Yoshiaki
AU - Mochizuki, Yasuhiro
AU - Tanaka, Toshiya
AU - Hamakubo, Takao
AU - Sakai, Juro
AU - Kodama, Tatsuhiko
AU - Inoue, Tsuyoshi
AU - Tachibana, Keisuke
AU - Doi, Takefumi
N1 - Publisher Copyright:
© 2016 Ishimoto et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2016/10
Y1 - 2016/10
N2 - Posttranslational modifications (PTMs) of proteins play a crucial role in regulating proteinprotein 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.
AB - Posttranslational modifications (PTMs) of proteins play a crucial role in regulating proteinprotein 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.
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U2 - 10.1371/journal.pone.0165766
DO - 10.1371/journal.pone.0165766
M3 - Article
C2 - 27798683
AN - SCOPUS:84994609049
VL - 11
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 10
M1 - e0165766
ER -