Innovative Drug Development for Histone Methylase

Research Project

Project/Area Number	25670026
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Biological pharmacy
Research Institution	Osaka University
Principal Investigator	DOI Takefumi 大阪大学, 薬学研究科(研究院), 教授 (00211409)
Project Period (FY)	2013-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000) Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	ヒストンH３ / ヒストンメチル化 / SETDB1 / MCAF1 / 翻訳後修飾 / ユビキチン化 / ヒストンH3 / ヒストンメチル化酵素 / タンパク質精製
Outline of Final Research Achievements	SETDB1 is one of the H3K9 methylation enzymes and requires MCAF1 for H3K9’s tri-methylation. In this study, we have investigated the interaction between SETDB1 and MCAF1 to elucidate the tri-methylation mechanism by SETDB1. We first confirmed that SETDB1(1-195) and MCAF1(562-817) formed a complex by immune-precipitation experiment. Then, in order to reveal the complex structure, we expressed these proteins in E.coli and purified them for further X-ray analysis. Next, we searched the relationship between the post-translational modification and methylation activity of SETDB1. Analyses of several deletion mutants checking their methylation activities and modification revealed that the ubiquitination of SETDB1 is needed for its methylation activity.