Revealing the physiological function of de-N-glycosylating enzymes in mice

• Project/Area Number: 16K18520
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Juntendo University
• Principal Investigator: Fujihira Haruhiko 順天堂大学, 医学部, 特任研究員 (50721057)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: Ngly1 / ENGase / ubiquitin-proteasome / autophagy / 糖鎖脱離酵素 / PNGase / ERAD / Autophagy / ユビキチン・プロテアソーム / オートファジー

Outline of Final Research Achievements

In this study, I aimed to reveal an unknown physiological function of de-N-glycosylating enzymes, peptide:N-glycanase (Ngly1) and endo-beta-N-acetylglucosaminidase (ENGase), in mammals. To this end, I performed phenotypic analyses of mice that lack Ngly1/Engase gene (Ngly1-KO, Engase-KO, Ngly1;Engase-KO mice). I found that the deletion of Ngly1 causes many defects (heart, muscle, nervous systems, body weight etc), while Engase deletion does not cause any defects. Interestingly, the defects caused by the loss of Ngly1 are partially rescued by the additional deletion of Engase gene. Therefore, there are two types of pathways that cause the defects by Ngly1-deletion, ENGase-dependent pathway and ENGase-independent pathway. Based on recent reports and my additional experiments suggested that a transcriptional factor Nrf1, which regulates the expression levels of proteasome subunits under low proteasome activity, seems to be involved in the ENGase-independent pathway.

Research Products

• [Journal Article] A Critical Domain of Ebolavirus Envelope Glycoprotein Determines Glycoform and Infectivity (2018)
  • Author(s): Fujihira H, Usami K, Matsuno K, Takeuchi H, Denda-Nagai K, Furukawa JI, Shinohara Y, Takada A, Kawaoka Y, Irimura T
  • Journal Title: Sci Rep Volume: 8 Issue: 1 Pages: 5495-5495
  • DOI: 10.1038/s41598-018-23357-8
  • Peer Reviewed / Open Access
• [Journal Article] 60年間の研究から得た教訓 (2017)
  • Author(s): 藤平陽彦、鈴木匡
  • Journal Title: 生化学 Volume: 89 Issue: 5 Pages: 599-599
  • DOI: 10.14952/SEIKAGAKU.2017.890599
  • ISSN: 0037-1017
  • Year and Date: 2017-10-25
  • Peer Reviewed
• [Journal Article] Lethality of mice bearing a knockout of the Ngly1-gene is rescued by the additional deletion of the Engase gene. (2017)
  • Author(s): Fujihira H., Masahara-Negishi Y., Tamura M., Harada Y., Chengcheng H., Wakana S., Taniguchi N., Kondoh G., Yamashita T., Funakoshi Y. and Suzuki T.
  • Journal Title: PLOS Genetics Volume: 13 Issue: 4 Pages: 1-23
  • DOI: 10.1371/journal.pgen.1006696
  • Peer Reviewed / Open Access
• [Presentation] The defects caused by the loss of Ngly1 can be partially rescued by the additional deletion of Engase (2018)
  • Author(s): Haruhiko Fujihira, Yuki Masahara-Negishi, Masaru Tamura, Shigeharu Wakana, Daisuke Takakura, Nana Kawasaki, Yoko Funakoshi, Tadashi Suzuki
  • Organizer: International Symposium on ER Stress, glycosylation, homeostasis and diseases
  • Int'l Joint Research
• [Presentation] Engase欠損により回避可能/不可能なNgly1-KOマウスの表現型 (2017)
  • Author(s): 藤平陽彦
  • Organizer: 第3回 ENGase研究会
• [Presentation] Summary of phenotypic analyses of Ngly1-KO mice and Ngly1/Engase-KO mice (2017)
  • Author(s): Haruhiko Fujihira, Yuki Masahara-Negishi, Masaru Tamura, Shigeharu Wakana, Daisuke Takakura, Nana Kawasaki, Yoko Funakoshi, Tadashi Suzuki
  • Organizer: International Syposium “Systems Glycobiology and Beyond”
  • Int'l Joint Research
• [Presentation] Engase欠損により回避可能/不可能なNgly1欠損による異常 (2017)
  • Author(s): 藤平陽彦、根岸-正原由紀、田村勝、高倉大輔、黄澄澄、若菜茂晴、川崎ナナ、船越陽子、鈴木匡
  • Organizer: 2017年度生命科学系学会合同年次大会(ConBio2017)
• [Presentation] Physiological function of deglycosylating enzymes acting on N-glycans in mice (2016)
  • Author(s): Haruhiko Fujihira, Yuki Masahara-Negishi, Masaru Tamura, Shigeharu Wakana, Daisuke Takakura, Nana Kawasaki, Yoko Funakoshi, Tadashi Suzuki
  • Organizer: Frontiers 2016 symposium
  • Place of Presentation: EPFL (Ecole Polytechnique Federale de Lausanne) (Switzerland)
  • Year and Date: 2016-12-05
  • Int'l Joint Research