DEVELOPMENT OF A CROSSLINKING-BASED ASSAY FOR CHROMATIN RESEARCH

• Project/Area Number: 22510219
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: System genome science
• Research Institution: Fujita Health University
• Principal Investigator: ISHIHARA Satoru 藤田保健衛生大学, 医学部, 講師 (00300723)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: ゲノム解析技術 / クロマチン / ヌクレオソーム / ホルマリン / 転写 / SEVENS法 / プロモーター

Research Abstract

The treatment of chromatin with formaldehyde makes nucleosomesto be cross-linked to each other. When there are more nucleosomes in the vicinity, thecross-linking reaction occurs well to produce larger particles including the nucleosomes.These particles can be fractionated in the sedimentation velocity centrifugation, inwhich nucleosomes cross-linked well are separated from ones cross-linked poorly. Here,the fractionation (designated as the SEVENS assay) was developed for chromatin researchapplicable to all mammalian cell types.

Research Products

• [Journal Article] The roleof IL-15 in activating STAT5 and fine- tuning IL-17A production in CD4 T lymphocytes (2012)
  • Author(s): Pandiyan,P., Yang,XP., Saravanamuthu, SS., Zheng,L., Ishihara,S., O'Shea, JJ., and Lenardo, MJ
  • Journal Title: J. Immunol Volume: 189 Issue: 9 Pages: 4237-4246
  • DOI: 10.4049/jimmunol.1201476
  • Peer Reviewed
• [Journal Article] Two-step binding of transcription factors causes sequential chromatin structural changes at the activated IL-2 promoter (2011)
  • Author(s): Ishihara, S. and Schwartz, RH
  • Journal Title: J. Immunol Volume: 187 Issue: 6 Pages: 3292-3299
  • DOI: 10.4049/jimmunol.1003173
  • Peer Reviewed
• [Journal Article] Two-step binding of transcription factors causes seguential chromatin structural changes at the activated IL-2 promoter (2011)
  • Author(s): Satoru Ishihara
  • Journal Title: The Journal of Immunology Volume: 187 Pages: 3292-3299
  • Peer Reviewed
• [Journal Article] A new fractionation assay,based on the size of formaldehyde crosslinked, mildly sheared chroma-tin, delineates the chromatin struc-ture at promoter regions (2010)
  • Author(s): Ishihara, S., Varma, R., and Schwartz,RH
  • Journal Title: Nucleic Acids Res Volume: 38 Issue: 11 Pages: e124-e124
  • DOI: 10.1093/nar/gkq203
  • Peer Reviewed
• [Journal Article] A new fractionation assay, based on the size of formaldehyde-crosslinked, mildly sheared chromatin, delineates the chromatin structure at promoter regions. (2010)
  • Author(s): Satoru Ishihara
  • Journal Title: Nucleic Acids Research Volume: 38
  • Peer Reviewed
• [Presentation] The chromatin structure responsible for the degree of the promoter activity (2012)
  • Author(s): 琴村直恵,原田信広,石原悟
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Presentation] CYP19遺伝子のプロモーター選択機構:密なクロマチン構造でプロモーターを選択的に抑制する (2012)
  • Author(s): 石原悟,琴村直恵,原田信広
  • Organizer: 第6回日本エピジェネティクス研究会年会
  • Place of Presentation: 東京.
• [Presentation] CYP19遺伝子のプロモーター選択機構：密なクロマチン構造でプロモーターを選択的に抑制する (2012)
  • Author(s): 石原悟、琴村直恵、原田信広
  • Organizer: 第6回日本エピジェネティクス研究会年会
  • Place of Presentation: 東京
• [Presentation] The chromatin structure responsible for the degree of the promoter activity. (2012)
  • Author(s): Kotomura, N., Harada, N., Ishihara, S.
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Presentation] The contribution of histone modifications to alternative promoter usage of the human CYP19 gene (2011)
  • Author(s): 石原悟
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市)
  • Year and Date: 2011-12-16
• [Presentation] ヒストンH3K27のトリメチル化によるアロマターゼ遺伝子プロモーターの選択的不活性化 (2011)
  • Author(s): 石原悟,琴村直恵,西芳寛,野村政壽,柳瀬敏彦,原田信広
  • Organizer: 第19回日本ステロイドホルモン学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2011-11-26
• [Presentation] ヒストンH3K27のトリメチル化によるアロマターゼ遺伝子プロモーターの選択的不活性化 (2011)
  • Author(s): 石原悟
  • Organizer: 第19回日本ステロイドホルモン学会学術集会
  • Place of Presentation: 福岡大学病院メディカルホール(福岡県福岡市)
  • Year and Date: 2011-11-26
• [Presentation] Condensed Chromatin Marked by K27-Trimethylated Histone H3 Determines Alternative Promoter Usage of the CYP19 Gene (2011)
  • Author(s): 石原悟
  • Organizer: 第5回日本エピジェネティクス研究会年会
  • Place of Presentation: KKRホテル熊本(熊本県熊本市)
  • Year and Date: 2011-05-19
• [Presentation] The contribution of histone modifica-tions to alternative promoter usage of the human CYP19 gene. (2011)
  • Author(s): 琴村直恵,原田信広,石原悟
  • Organizer: 第34回日本分子生物学会年会.
  • Place of Presentation: 横浜
• [Presentation] CondensedChromatin Marked by K27-Trimethy-lated Histone H3 Determines Alterna-tive Promoter Usage of the CYP19 Gene (2011)
  • Author(s): 石原悟,琴村直恵,原田信広
  • Organizer: 第5回日本エピジェネティクス研究会年会
  • Place of Presentation: 熊本
• [Presentation] Condensed chromatin marked by K27-trimethylated histone H3 determines alternative promoter usage of the CYP19 gene (2011)
  • Author(s): Kotomura,N.,Harada,N.,Ishihara,S
  • Organizer: CDB Symposium"Epigene- tic landscape in development and dis-ease
  • Place of Presentation: Kobe,JAPAN
• [Presentation] Sequential rearrangement and eviction of nucleo-somes allow interleukin-2 transcrip-tion following T cell activation. (2010)
  • Author(s): Ishihara,S.,Schwartz,RH
  • Organizer: The NIBB Conference "DYNAMICGENOME
  • Place of Presentation: Okazaki, JAPAN
  • Year and Date: 2010-10-16
• [Presentation] Sequential rearrangement and eviction of nucleosomes allow interleukin-2 transcription following T cell activation. (2010)
  • Author(s): 石原悟
  • Organizer: The NIBB Conference "DYNAMIC GENOME"
  • Place of Presentation: 自然科学研究機構岡崎コンファレンスセンター(愛知県岡崎市)
  • Year and Date: 2010-10-16