Mechanism of ovarian carcinogenesis due to 26S proteasome activation by protooncogene gankyrin mononeddylation

• Project/Area Number: 24592514
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: Kyoto University
• Principal Investigator: HIGASHITSUJI Hisako 京都大学, 医学(系)研究科(研究院), 助教 (20402852)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: gankyrin / mononeddylation / S6 ATPase / 26S proteasome / polyubiquitylation / synthetic biology / mdm2 / p53 / NEDD8 / E3 ubiquitin ligase / pRB / 卵巣がん / ガンキリン / プロテアソーム / 卵巣癌 / 癌遺伝子

Outline of Final Research Achievements

In ovarian cancers, gankyrin and Nedd8 are overexpressed. In vitro gankyrin neddylation assay (bacteria-expressed and purified E1, E2, Mdm2, Nedd8, gankyrin, ATP) produces gankyrin mononeddylation, that is verified by western blot of anti-gankyrin antibody and anti-Nedd8 antibody. Due to synthetic biological method, polycistronic affinity-tagged gankyrin, Nedd8, E1, E2, and Mdm2 are expressed into Rosetta bacterial cells. Gankyrin mononeddylation occurs. This expressed and purified mononeddylated gankyrin enhances p53 and pRB polyubiquitylation and 26S proteasome-mediated protein degradation. This effect is enhanced by full-length S6 ATPase co-expression. However it is suppressed by C-terminal truncated S6 ATPase mutant.

Research Products

• [Journal Article] Stress response protein cirp links inflammation and tumorigenesis in colitis-associated cancer. (2014)
  • Author(s): Sakurai T, Kashida H, Watanabe T, Hagiwara S, Mizushima T, Iijima H, Nishida N, Higashitsuji H, Fujita J, Kudo M.
  • Journal Title: Cancer Res. Volume: 74 Pages: 6119-28
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Overexpression of gankyrin in mouse hepatocytes induces hemangioma by suppressing factor inhibiting hypoxia-inducible factor-1 (FIH-1) and activating hypoxia-inducible factor-1. (2014)
  • Author(s): Liu Y, Higashitsuji H, Higashitsuji H, Itoh K, Sakurai T, Koike K, Hirota K, Fukumoto M, Fujita J.
  • Journal Title: Biochem Biophys Res Commun. Volume: 432(1) Pages: 22-27
  • Peer Reviewed
• [Journal Article] Hypothermia protects against fulminant hepatitis in mice by reducing reactive oxygen species production. (2013)
  • Author(s): Sakurai T, Kudo M, Watanabe T, Itoh K, Higashitsuji H, Arizumi T, Inoue T, Hagiwara S, Ueshima K, Nishida N, Fukumoto M, Fujita J.
  • Journal Title: Dig Dis Volume: 31 Issue: 5-6 Pages: 440-6
  • DOI: 10.1159/000355242
  • Peer Reviewed
• [Journal Article] Identification of a novel enhancer that binds Sp1 and contributes to induction of cold-inducible RNA-binding protein (cirp) expression in mammalian cells. (2012)
  • Author(s): Sumitomo Y, Higashitsuji H, Higashitsuji H, Liu Y, Fujita T, Sakurai T, et al
  • Journal Title: BMC Biotechnol Volume: 12 Issue: 1 Pages: 72-72
  • DOI: 10.1186/1472-6750-12-72
  • Peer Reviewed
• [Journal Article] Cold-inducible RNA-binding protein (Cirp) interacts with Dyrk1b/Mirk and promotes proliferation of immature male germ cells in mice (2012)
  • Author(s): Masuda T, et al.
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Issue: 27 Pages: 10885-108901
  • DOI: 10.1073/pnas.1121524109
  • Peer Reviewed
• [Presentation] ほ乳類低温ショックタンパク質CIRPはBRCA1による相同組み替えによるDNA修復を促進する (2014)
  • Author(s): 東辻久子
  • Organizer: 第６９回日本消化器外科学会総会
  • Place of Presentation: 郡山市立文化センター
  • Year and Date: 2014-07-16 – 2014-07-18
• [Presentation] ほ乳類低温ショックタンパク質Cirpは皮膚創傷治癒を促進する (2013)
  • Author(s): 藤田潤、東辻久子、東辻宏明
  • Organizer: 第８回臨床ストレス応答学会
  • Place of Presentation: 松本市、信州大学医学部付属病院、大会議室
• [Presentation] Overexpression of gankyrin in mouse hepatocytes induces hemangioma. (2013)
  • Author(s): 東辻宏明、東辻久子、藤田潤
  • Organizer: 第２１回JDDW２０１３
  • Place of Presentation: 東京都品川区
• [Presentation] ACC1脂質代謝系酵素によるBRCA1のhomologous recombination の調節機序 (2013)
  • Author(s): 東辻宏明、東辻久子
  • Organizer: 日本消化器病学会
  • Place of Presentation: 鹿児島市