Functional analysis of the FANCD2 interactors containing ubiquitin binding domain

2013 Fiscal Year Final Research Report

• Project/Area Number: 23590380
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Human genetics
• Research Institution: Kyoto University
• Principal Investigator: ISHIAI Masamichi 京都大学, 放射線生物研究センター, 准教授 (90298844)
• Project Period (FY): 2011 – 2013
• Keywords: ユビキチン / DNA修復 / DNA損傷応答 / ファンコニ貧血 / 癌 / 環境 / FANCD2 / DT40細胞

Research Abstract

The Fanconi anemia (FA) pathway is critically involved in DNA damage responses and DNA repair. A key protein, FANCD2, is monoubiquitinated and accumulates in chromatin in response to DNA interstrand crosslinks (ICLs), where it coordinates DNA repair through mechanisms that are still poorly understand.
We have isolated UBR5 and UFD1 as candidate interacting proteins for FANCD2. Both proteins contain ubiquitin-binding domains. We have established ubr5-deficient chicken DT40 cells, and SiRNA-based ufd1-knock down human cells. Unexpectedly, ubr5-deficient or ufd1-knock down cells showed little sensitivity to ICL reagents compared to wild type cells. Laser microirradiation experiments using GFP-UBR5 or GFP-UFD1 expressing human cells showed little GFP-protein accumulation after DNA damage. These results suggested that functional relationship between FA pathway and UBR5, or UFD1 including related proteins was not strong, at least in ICL repair.

Research Products

• [Journal Article] FANCD2 binds CtIP and regulates DNA-end resection during DNA interstrand crosslink repair (2014)
  • Author(s): Unno, J., Itaya, A., Taoka, M., Sato, K., Tomida, J., Sakai, W., Sugasawa, K., Ishiai, M., Ikura, T., Isobe, T., Kurumizaka, H. Takata M
  • Journal Title: Cell Report Volume: 7 Pages: 1039-1047
  • DOI: 10.1016/j.celrep.2014.04.005
  • Peer Reviewed
• [Journal Article] Tumor suppressor RecQL5 controls recombination induced by DNA crosslinking agents (2014)
  • Author(s): Hosono, Y., Abe, T., Ishiai, M., Islam, M. N., Arakawa, H., Wang, W., Takeda, S., Ishii, Y., Takata, M., Seki, M. Enomoto T
  • Journal Title: Biochim Biophys Acta. Volume: 1843 Pages: 1002-1012
  • DOI: 10.1016/j.bbamcr.2014.01.005
  • Peer Reviewed
• [Journal Article] Histone chaperon activity of Fanconi anemia proteins, FANCD2 and FANCI, is required for DNA crosslink repair (2012)
  • Author(s): Sato, K., Ishiai, M., Toda, K., Furukoshi, S., Osakabe, A., Tachiwara, H., Takizawa, Y., Kagawa, W., Kitao, H., Dohmae, N., Obuse, C., Kimura H., Takata, M. and Kurumizaka H
  • Journal Title: EMBO J. Volume: 31 Pages: 3524-3536
  • DOI: 10.1038/emboj.2012.197
  • Peer Reviewed
• [Journal Article] Mcm8 and Mcm9 form a complex that functions in homologous recombination repair induced by DNA interstrand crosslinks (2012)
  • Author(s): Nishimura, K., Ishiai, M., Hirikawa, K., Fukagawa, T., Takata, M., Takisawa, H. Kanemaki, M.T
  • Journal Title: Mol. Cell Pages: 47
  • DOI: 10.1016/j.molcel.2012.05047
  • Peer Reviewed
• [Presentation] ファンコニ貧血原因遺伝子産物FANCD2の示すヒストンシャペロン活性の生理的意義 (2013)
  • Author(s): 石合正道, 佐藤浩一, 胡桃坂仁志, 高田穣
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20131203-06
  • Invited
• [Presentation] ファンコニ貧血経路の示すヌクレオソーム形成活性のDNA修復における役割 (2012)
  • Author(s): 石合正道, 佐藤浩一, 高田穣, 胡桃坂仁志
  • Organizer: 日本放射線影響学会第55回大会
  • Place of Presentation: 仙台市
  • Year and Date: 20120906-08
  • Invited
• [Presentation] Histone chaperon activity of FANCD2 Plays a crucial role in DNA crosslink repair (2012)
  • Author(s): Ishiai, M.,Sato, K., Toda, K., Furukoshi, S., Osakabe, A., Tachiwara, H., Takizawa, Y., Kagawa, W., Kitao, H., Dohmae, N., Obuse, C., Kimura H., Takata, M., Kurumizaka H
  • Organizer: Abcam International meeting on"Maintenance of Genome Stability 2012"
  • Place of Presentation: ナッソー市(バハマ国)
  • Year and Date: 20120305-08
• [Presentation] Molecular mechanisms of the Fanconi anemia pathway (2011)
  • Author(s): 石合正道, 冨田純也, 茂地智子, 板谷亜希子, 内田恵美, 海野純也, 高田穣
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜市
  • Year and Date: 20111213-16
  • Invited
• [Book] Establishiment of the DNA repair-defective mutants in DT40 cells in"DNA repair Protocols, Methods in Molecular Biology, vol. 920 (Bjerbak ed.)"(Chapter 4) (2012)
  • Author(s): Ishiai, M., Uchida, E., Takata, M.
  • Total Pages: 39-49
  • Publisher: Humana Press
• [Remarks]
  • URL: http://house.rbc.kyoto-u.ac.jp/late-effect