Study of human HLTF, a ubiquitin ligase for PCNA under DNA damage

• Project/Area Number: 23770125
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Yokohama City University
• Principal Investigator: HISHIKI Asami 横浜市立大学, 大学院・生命ナノシステム科学研究科, 特任助教 (60571172)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: X線結晶解析 / 国際情報交流

Research Abstract

Helicase-Like Transcription Factor (HLTF) is involved in template switching pathway, one of DNA damage tolerance mechanism, which allows continuous DNA synthesis even in the presence of DNA lesion. HLTF consists of RING, Snf2, and HIRAN domains. RING and Snf2 domains are responsible for ubiquitination of PCNA and unwinding of DNA, respectively. The N-terminal HIRAN domain is predicted as a novel DNA-binding domain. We tried to prepare human HLTF (hHLTF) RING and HIRAN domains as recombinant proteins. We analyzed interaction between hHLTF HIRAN domain and DNA by Electrophoresis Mobility Shift Assay.

Research Products

• [Journal Article] En bloc transfer of polyubiquitin chains to PCNA in vitro is mediated by two different human E2-E3 pairs (2012)
  • Author(s): Masuda Y
  • Journal Title: Nucleic Acids Res. Volume: 40(20) Issue: 20 Pages: 10394-10407
  • DOI: 10.1093/nar/gks763
  • Peer Reviewed
• [Journal Article] Identification of a small molecule PCNA inhibitor that disrupts interactions with PIP-Box proteins and inhibits DNA replication (2012)
  • Author(s): Chandanamali Punchihewa, Akira Inoue, Asami Hishiki, Yoshihiro Fujikawa, Michele Connelly, Benjamin Evison, Youming Shao, Richard Heath, Isao Kuraoka, Patrick Rodrigues, Hiroshi Hashimoto, Masanobu Kawanishi, Mamoru Sato, Takashi Yagi, Naoaki Fujii
  • Journal Title: J Biol Chem Volume: 287 Issue: 17 Pages: 14289-14300
  • DOI: 10.1074/jbc.m112.353201
  • Peer Reviewed
• [Presentation] ヒト PCNA のポリユビキチン化の新規分子機構 (2012)
  • Author(s): 増田雄司, 鈴木美紀, 河合秀彦, 菱木麻美, 橋本博, 益谷央豪, 神谷研二
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場
• [Presentation] Crystal structures of human PCNA in complex with small molecule inhibitors for cancer therapeutics (2012)
  • Author(s): Hashimoto H, Kikuchi S, Hishiki A, Punchihewa C, Fujii N, Sato M
  • Organizer: AsCA 12/CRYSTAL 28
  • Place of Presentation: Aderaide Convention Centre
• [Presentation] En bloc transfer of poly-ubiquitin chains to PCNA in vitro is mediated by two different human E2-E3 pairs (2012)
  • Author(s): Masuda Y, Suzuki M, Kawai H, Hishiki A, Hashimoto H, Masutani C, Hishida T, Suzuki F, Kamiya K
  • Organizer: The 8th 3R Symposium
  • Place of Presentation: Awaji Yumebutai International Conference Center
• [Presentation] 抗がん剤併用薬開発に向けた PCNAと阻害剤との複合体構造 (2012)
  • Author(s): 菊池壮太郎, 菱木麻美, Chandanamali Punchihewa, 藤井直明, 佐藤衛, 橋本博
  • Organizer: 第12回日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場
• [Presentation] ヒトPCNAのポリユビキチン化の新規分子機構 (2012)
  • Author(s): 増田雄司, 鈴木美紀, 河合秀彦, 菱木麻美, 橋本 博, 益谷央豪, 神谷研二
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場
• [Presentation] En bloc transfer of poly-ubiquitin chains to PCNA in vitro is mediated by two different human E2-E3 pairs. (2012)
  • Author(s): Masuda, Y., Suzuki, M., Kawai, H., Hishiki, A., Hashimoto, H., Masutani, C., Hishida, T., Suzuki, F., Kamiya, K.
  • Organizer: The 8th 3R Symposium
  • Place of Presentation: Awaji Yumebutai International Conference Center
• [Presentation] 抗がん剤併用薬開発に向けたPCNAと阻害剤との複合体構造 (2012)
  • Author(s): 菊池壮太郎、菱木麻美、Chandanamali Punchihewa、藤井直明、佐藤衛、橋本博
  • Organizer: 第12回日本蛋白質科学会年会
  • Place of Presentation: 第12回日本蛋白質科学会年会