Crystallographic analysis of the FACT-histone complex

2010 Fiscal Year Final Research Report

• Project/Area Number: 20370043
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: SENDA Toshiya National Institute of Advanced Industrial Science and Technology, バイオメディシナル情報研究センター, 主任研究員 (30272868)
• Co-Investigator(Renkei-kenkyūsha): HORIKOSHI Masami 東京大学, 分子細胞生物学研究所, 准教授 (70242089) ; ENOMOTO Takemi 武蔵野大学, 薬学研究所, 教授 (80107383)
• Project Period (FY): 2008 – 2010
• Keywords: 結晶構造解析 / 結晶化 / ヒストン / ヌクレオソーム / ヒストンシャペロン / 精製

Research Abstract

FACT, which interacts with the histone H2A-H2B complex, is known to be involved in nucleosome assembly/disassembly. In order to reveal the molecular mechanism of nucleosome assembly/disassembly, crystal structure analysis of FACT was initiated. In the present study, we have established a method that can give about 7 mg of highly purified FACT from 1L culture of insect cell SF+. In addition, a purification method of histone proteins was improved. We confirmed that the purified FACT interacts with the histone proteins. To establish a proper condition for preparing the FACT-histone complex without forming aggregates, solution conditions of the FACT-histone complex were investigated by the dynamic light scattering method. On the basis these results, we are making an effort to obtain crystals of the FACT-histone complex.

Research Products

• [Journal Article] Structure of the histone chaperone CIA/ASF1-double bromodomain complex linking histone modifications and site-specific histone eviction. (2010)
  • Author(s): Akai, Y., Adachi, N., Hayashi, Y., Eitoku, M., Sano, N., Natsume, R., Kudo, N., Tanokura, M., Senda, T. Horikoshi, M.
  • Journal Title: Proc.Natl.Acad.Sci.USA. 107 Pages: 8153-8158
  • Peer Reviewed
• [Journal Article] Theoretical framework for the histone modification network : modifications in the unstructured histone tails form a scale-free network. (2009)
  • Author(s): Hayashi, Y., Senda, T., Sano, N. Horikoshi, M.
  • Journal Title: Genes Cells 14 Pages: 789-806
  • Peer Reviewed
• [Journal Article] Crystal structure of Methanococcus jannaschii TATA box-binding protein. (2008)
  • Author(s): Adachi, N.,Senda, M., Natsume, R., Senda, T., Horikoshi, M.
  • Journal Title: Genes Cells 13 Pages: 1127-1140
  • Peer Reviewed
• [Journal Article] Effect of leucine-to-methionine substitutions on the diffraction quality of histone chaperone SET/TAF-Ibeta/INHAT crystals. (2008)
  • Author(s): Senda, M., Muto, S., Horikoshi, M., Senda, T.
  • Journal Title: Acta Crystallogr.F 64 Pages: 960-965
  • Peer Reviewed
• [Presentation] Structure-Based Model of the Biological Signaling from Histone Modifications to Structural Change of the Nucleosom (2011)
  • Author(s): Senda, T.
  • Organizer: International symposium on the physicochemical field for genetic activities
  • Place of Presentation: Awaji, Japan
  • Year and Date: 2011-01-25
• [Presentation] ヒストンシャペロン複合体の結晶構造解析 (2010)
  • Author(s): 千田俊哉
  • Organizer: BMB2010
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-10
• [Presentation] CIA/ASF1を中心としたヌクレオソーム構造変換機構 (2010)
  • Author(s): 千田俊哉
  • Organizer: 構造エピゲノム研究会発足記念シンポジウム
  • Place of Presentation: 横浜
  • Year and Date: 2010-04-30
• [Presentation] Molecular mechanism of the site-specific histone eviction at active promoter regions (2009)
  • Author(s): Senda, T.
  • Organizer: Taiwan-Japan Symposium on crystallography.
  • Place of Presentation: Osaka Japan
  • Year and Date: 2009-12-08
• [Presentation] Structure and function of the human histone chaperone CIA complexed with the bromodomain from TFIID (2008)
  • Author(s): Akai, Y.
  • Organizer: XXI congress and general assembly of the in ternational union of crystallography.
  • Place of Presentation: Osaka Japan
  • Year and Date: 2008-08-28
• [Presentation] Structure and function of the histone chaperone CIA/ASF1 complexed with histone H3 and H4 (2008)
  • Author(s): Natsume, R.
  • Organizer: XXI congress and general assembly of the international union of crystallography.
  • Place of Presentation: Osaka Japan
  • Year and Date: 2008-08-26