Elucidating the mechanisms underlying linear ubiquitin chains-mediated cell death protection

• Project/Area Number: 15K19007
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: Fujita Hiroaki 京都大学, 医学研究科, 助教 (90738006)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 細胞死 / LUBAC / 直鎖状ポリユビキチン鎖

Outline of Final Research Achievements

LUBAC is involved in TNF-alpha mediated NF-kappaB activation via generating linear polyubiquitin chains. We discovered that LUBAC is also involved in the protection of TNF-alpha-mediated cell death in NF-kappaB independent manner. To elucidate the mechanisms, I prepared LUBAC knockout cells and reconstituted the several LUBAC mutants into cells to analyze whose domains are involved in cell death protection. Finally, we found a domain of LUBAC which participates in cell death regulation and explored proteins which bind to newly discovered domain by Mass spectrometry.

Research Products

• [Journal Article] Differential involvement of the NZF domains of SHARPIN and HOIL-1L in LUBAC-mediated cell death protection. (2016)
  • Author(s): Shimizu S, Fujita H, Sasaki Y, Tsuruyama T, Fukuda K, Iwai K.
  • Journal Title: Mol Cell Biol. Volume: 01049-15 Issue: 10 Pages: 1569-1583
  • DOI: 10.1128/mcb.01049-15
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Crystal structure of the N-myristoylated lipopeptide-bound MHC class I complex. (2015)
  • Author(s): Morita D, Yamamoto Y, Mizutani T, Ishikawa T, Suzuki J, Igarashi T, Mori N, Shiina T, Inoko H, Fujita H, Iwai K, Tanaka Y, Mikami B, Sugita M,
  • Journal Title: Nat Commun. Volume: 7 Issue: 1 Pages: 10356-10356
  • DOI: 10.1038/ncomms10356
  • NAID: 120005743661
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Immunoblot Analysis of Linear Polyubiquitination of NEMO. (2015)
  • Author(s): Sasaki, Y., Fujita, H., Nakai, M., and Iwai, K.
  • Journal Title: Methods in Molecular Biology, NF-kappaB Methods and Protocols Volume: 1280 Pages: 297-309
  • DOI: 10.1007/978-1-4939-2422-6_17
  • ISBN: 9781493924219, 9781493924226
  • Acknowledgement Compliant
• [Presentation] Novel insights into linear ubiquitin chain assembly complex formation (2016)
  • Author(s): 藤田宏明
  • Organizer: KEYSTONE SYMPOSIA
  • Place of Presentation: Whistler, British Columbia, Canada
  • Year and Date: 2016-03-13
  • Int'l Joint Research
• [Presentation] Crucial roles of unexpected interaction between subunits of LUBAC ubiquitin ligase in NF-kappaB and cell death (2016)
  • Author(s): Hiroaki F., Satoshi S., Kazuhiro I.
  • Organizer: 第39回日本分子生物学会 シンポジウム 口頭発表
  • Place of Presentation: パシフィコ横浜
• [Presentation] LUBACリガーゼ形成における相同性の高いHOIL-1L、SHARPINのUBLの機能的差異 (2015)
  • Author(s): 藤田宏明
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] LUBAC安定化機構の解析からみえてきたHOIL-1L、SHARPINの機能的差異 (2015)
  • Author(s): 藤田宏明
  • Organizer: シグナルネットワーク研究会
  • Place of Presentation: 沖縄科学技術大学院大学
  • Year and Date: 2015-06-19
• [Book] シグナリング研究2015 (2015)
  • Author(s): 藤田宏明、岩井一宏
  • Total Pages: 7
  • Publisher: 羊土社