Deciphering the factors associated with selective demethylation of tissue-specific gene promoter

• Project/Area Number: 16K21142
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biological pharmacy; Applied molecular and cellular biology
• Research Institution: Osaka University
• Principal Investigator: Hino Nobumasa 大阪大学, 薬学研究科, 助教 (90469916)
• Research Collaborator: TAKASHIMA Seiji 大阪大学, 大学院生命機能研究科, 教授 ; OKADA Yoshiaki 大阪大学, 大学院薬学研究科, 准教授 ; KITA Ayami 大阪大学, 大学院薬学研究科, 大学院生
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: エピジェネティクス / 組織特異的遺伝子 / 脱メチル化 / タンパク質間相互作用 / 光クロスリンク / 非天然型アミノ酸 / クロスリンク / 人工アミノ酸 / DNA脱メチル化

Outline of Final Research Achievements

Expression of tissue-specific gene is regulated by DNA methylation of its promoter region. The hypermethylated promoter is demethylated along with differentiation of the cells to induce the downstream gene expression. The demethylation is regulated by TET family enzymes (TET1-3), however, it is not well understood how these enzymes recognize promoters that should be demethylated at specific time point during cell differentiation. In this study, we attempted to identify TET1-interacting proteins that can recruit TET1 to specific promoter regions. We employed in vivo protein photo-cross-linking technique and identified a novel protein that binds to TET1 catalytic domain in living cells. Interestingly, the active form of this protein directly promoted the enzymatic activity of TET1 in vitro. Considered with the fact that this protein is activated during differentiation of several cell types, our findings suggest the existence of TET1-activity modulating mechanism at cell differentiation.

Research Products

• [Journal Article] ETV2-TET1/TET2 Complexes Induce Endothelial Cell-Specific Robo4 Expression via Promoter Demethylation. (2018)
  • Author(s): Tanaka Toru、Izawa Kohei、Maniwa Yusuke、Okamura Maki、Okada Atsumasa、Yamaguchi Tomoko、Shirakura Keisuke、Maekawa Naoki、Matsui Hayato、Ishimoto Kenji、Hino Nobumasa、Nakagawa Osamu、Aird William C.、Mizuguchi Hiroyuki、Kawabata Kenji、Doi Takefumi、Okada Yoshiaki
  • Journal Title: Sci Rep. Volume: 8(1) Issue: 1 Pages: 5653-5653
  • DOI: 10.1038/s41598-018-23937-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Endothelial Robo4 regulates IL-6 production by endothelial cells and monocytes via a crosstalk mechanism in inflammation (2018)
  • Author(s): Shirakura Keisuke、Ishiba Ryosuke、Kashio Taito、Sakai Miki、Fukushima Yu、Yamamoto Nana、Manabe Shiori、Shigesada Naoya、Tanaka Toru、Hino Nobumasa、Aird William C.、Doi Takefumi、Okada Yoshiaki
  • Journal Title: Biochem Biophys Res Commun. Volume: 495 Issue: 1 Pages: 801-806
  • DOI: 10.1016/j.bbrc.2017.11.067
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Degradation of human Lipin-1 by BTRC E3 ubiquitin ligase (2017)
  • Author(s): Ishimoto Kenji、Hayase Ayaka、Kumagai Fumiko、Kawai Megumi、Okuno Hiroko、Hino Nobumasa、Okada Yoshiaki、Kawamura Takeshi、Tanaka Toshiya、Hamakubo Takao、Sakai Juro、Kodama Tatsuhiko、Tachibana Keisuke、Doi Takefumi
  • Journal Title: Biochem Biophys Res Commun. Volume: 488 Issue: 1 Pages: 159-164
  • DOI: 10.1016/j.bbrc.2017.04.159
  • Peer Reviewed
• [Journal Article] Tumor Necrosis Factor α Induces the Expression of the Endothelial Cell-Specific Receptor Roundabout4 through the Nuclear Factor-κB Pathway (2017)
  • Author(s): Tanaka T, Maekawa N, Kashio T, Izawa K, Ishiba R, Shirakura K, Ishimoto K, Hino N, Aird WC, Doi T, Okada Y.
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 40 Issue: 4 Pages: 504-509
  • DOI: 10.1248/bpb.b16-00938
  • NAID: 130005530067
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Adenovirus vector-based incorporation of a photo-cross-linkable amino acid into proteins in human primary cells and cancerous cell lines. (2016)
  • Author(s): Kita A, Hino N, Higashi S, Hirota K, Narumi R, Adachi J, Takafuji K, Ishimoto K, Okada Y, Sakamoto K, Tomonaga T, Takashima S, Mizuguchi H, Doi T.
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 36946-36946
  • DOI: 10.1038/srep36946
  • Peer Reviewed / Open Access
• [Journal Article] Ubiquitination of Lysine 867 of the Human SETDB1 Protein Upregulates Its Histone H3 Lysine 9 (H3K9) Methyltransferase Activity. (2016)
  • Author(s): Ishimoto K, Kawamata N, Uchihara Y, Okubo M, Fujimoto R, Gotoh E, Kakinouchi K, Mizohata E, Hino N, Okada Y, Mochizuki Y, Tanaka T, Hamakubo T, Sakai J, Kodama T, Inoue T, Tachibana K, Doi T
  • Journal Title: PLoS One Volume: 11 Issue: 10 Pages: e0165766-e0165766
  • DOI: 10.1371/journal.pone.0165766
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 細胞内光クロスリンク法によるTET1相互作用因子の探索 (2017)
  • Author(s): 喜多絢海、東咲子、Jeremia Febrian、高島成二、樋野展正、土井健史
  • Organizer: 日本薬学会 第137年会
  • Place of Presentation: 東北大学
  • Year and Date: 2017-03-24
• [Presentation] がん変異集積表面への光クロスリンカー導入による新規KEAP1結合因子の同定 (2017)
  • Author(s): 樋野展正，廣田康二，鳴海良平，堀越亮太，向山樹，足立淳，朝長毅， 土井健史
  • Organizer: 日本ケミカルバイオロジー学会
• [Presentation] 細胞内光クロスリンク法によるがん変異特異的タンパク質間相互作用解析 (2017)
  • Author(s): 廣田康二，樋野展正，鳴海良平，堀越亮太，向山樹，足立淳，朝長毅， 土井健史
  • Organizer: 日本分子生物学会
• [Presentation] アデノウイルスベクターによる光架橋性アミノ酸のタンパク質への導入 (2016)
  • Author(s): 喜多絢海、樋野展正、東咲子、廣田康二、高藤和輝、高島成二、水口裕之、土井健史
  • Organizer: 第15回 次世代を担う若手ファーマ・バイオフォーラム2016
  • Place of Presentation: 大阪大学
  • Year and Date: 2016-09-10
• [Book] Covalently Capturing Protein Interactions in Living Cells by Site-Specific Incorporation of Photo-Cross-Linkable Amino Acids. In: Hatanaka Y., Hashimoto M. (eds) Photoaffinity Labeling for Structural Probing Within Protein. (2017)
  • Author(s): Nobumasa Hino, Kensaku Sakamoto
  • Total Pages: 265
  • Publisher: Springer