Insight into how severely misfolded proteins are degraded in the ER

• Project/Area Number: 16K18538
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: Kyoto University
• Principal Investigator: Ninagawa Satoshi 京都大学, 理学研究科, 特定助教 (80647991)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: タンパク質分解 / 糖鎖 / 小胞体 / 構造異常タンパク質 / 小胞体関連分解 / mannose trimming / EDEM

Outline of Final Research Achievements

ER-associated degradation (ERAD) is composed of glycoprotein and non-glycoprotein degradation pathways. By inhibition of glycoprotein degradation pathway, native but unstable or somewhat unfolded glycoproteins were stabilized, but degradation of severely misfolded glycoproteins was delayed only at early chase periods, but they were eventually degraded as in wild-type cells. We analyzed these molecular mechanisms.
We tried to knocked out the candidate gene, which target substrates from glycoprotein degradation pathway to non-glycoprotein degradation pathway, and found that the gene is essential for cells, indicating its importance. Using PITCh method, its low expression strains were established. The mutant strain includes some mutations in genomic level. We found that the strain is defective to degrade severely misfolded non-glycoproteins.

Research Products

• [Journal Article] SEL1L-dependent Substrates Require Derlin2/3 and Herp1/2 for Endoplasmic Reticulum-associated Degradation (2017)
  • Author(s): Takehiro Sugimoto, Satoshi Ninagawa, Shimpei Yamano, Tokiro Ishikawa, Tetsuya Okada, Shunichi Takeda, Kazutoshi Mori
  • Journal Title: Cell Structure and Function Volume: 42 Issue: 2 Pages: 81-94
  • DOI: 10.1247/csf.17007
  • NAID: 130005708193
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access
• [Journal Article] UPR Transducer BBF2H7 Allows Export of Type II Collagen in a Cargo- and Developmental Stage-Specific Manner (2017)
  • Author(s): Tokiro Ishikawa, Takuya Toyama, Yuki Nakamura, Kentaro Tamada, Hitomi Shimizu, Satoshi Ninagawa, Tetsuya Okada, Yasuhiro Kamei, Tomoko Ishikawa-Fujiwara, Takeshi Todo, Eriko Aoyama, Masaharu Takigawa, Akihiro Harada and Kazutoshi Mori
  • Journal Title: Journal of cell biology Volume: 印刷中 Issue: 6 Pages: 1761-1774
  • DOI: 10.1083/jcb.201609100
  • NAID: 120006343784
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Direct mapping of additional modifications on phosphorylated O-glycans of α-dystroglycan by mass spectrometry analysis in conjunction with knocking out of causative genes for dystroglycanopathy (2016)
  • Author(s): H.Yagi, C.-W.Kuo, T.Obayashi, S.Ninagawa, K.-H.Khoo, and K.Kato
  • Journal Title: Mol. Cell Proteomics Volume: 15 Issue: 11 Pages: 3424-3434
  • DOI: 10.1074/mcp.m116.062729
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] 第二世代抗精神病薬オランザピンが、副作用として糖尿病を引き起こす分子メカニズムの解析 ～インスリンは、適切なジスルフィド結合形成を阻害され、小胞体関連分解により処理される～ (2017)
  • Author(s): Satoshi Ninagawa, Tetsuya Okada, Hajime Umezawa, Tokiro Ishikawa, Seiji Torii, Robert B. Mackin, Takahiko Nagamine, and Kazutoshi Mori
  • Organizer: ConBio2017
  • Int'l Joint Research