NF-YA欠損によるユビキチン蓄積病態の解析

• Project Area: Prevention of brain protein aging and dementia
• Project/Area Number: 15H01567
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Doshisha University
• Principal Investigator: 貫名 信行 同志社大学, 脳科学研究科, 教授 (10134595)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2016: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: NF-Y / 転写因子 / シャペロン / 神経変性 / ユビキチン / ポリグルタミン病 / 老化

Outline of Annual Research Achievements

転写因子NF-Yは、ハンチントン病、球脊髄性筋萎縮症などのポリグルタミン病モデルにおいて、様々な中枢神経系領域で機能阻害されることが見出されてきた。しかしながら、異なる神経細胞種での機能阻害が同様の変性病態を引き起こすかについては不明であった。
転写因子NF-YAに関してシャペロン系との関係を報告(EMBO J2008)するとともに、最近神経細胞においてノックアウトすることによりユビキチン、p62の集積を認めながら、線維性病変を伴わず、小胞体膜、核膜異常を伴う新規病変を見出した(Nat Comm2014)。今回我々は、NF-Yのノックダウンが線条体や小脳では大脳皮質と同様にユビキチン・p62蓄積病態を示すのに対し、脊髄運動神経細胞ではこれらのタンパク質蓄積を示さないことを見出した。運動神経細胞では前者の神経細胞群とは異なり、小胞体シャペロンGrp78（Bip）の発現が抑制されていた。Grp78の選択的発現抑制が、神経細胞種に特異的な変性病態に関わっていることが示唆された。本研究は細胞の異なる反応性が細胞特異的変性のメカニズムに関与していることを示唆している。（Yamanaka et al. Sci Rep 2016）。
一方、大脳皮質でのユビキチン蓄積病態のメカニズムを解明すべく生化学的解析を進め、プロテアーゼ処理によるいわゆる「蓄積コアタンパク質」の同定を試み、Proteinase K 処理抵抗性のユビキチン抗体に染まるバンドを見出した。この成分を質量分析により解析し、いくつかの候補タンパク質を同定した。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Aggregation of scaffolding protein DISC1 dysregulates phosphodiesterase 4 in Huntington's disease. (2017)
  • Author(s): Tanaka M, Ishizuka K, Nekooki-Machida Y, Endo R, Takashima N, Sasaki H, Komi Y, Gathercole A, Huston E, Ishii K, Hui KK, Kurosawa M, Kim SH, Nukina N, Takimoto E, Houslay MD, Sawa A.
  • Journal Title: J Clin Invest. Volume: 127 Issue: 4 Pages: 1438-1450
  • DOI: 10.1172/jci85594
  • Peer Reviewed
• [Journal Article] FUS/TLS acts as an aggregation-dependent modifier of polyglutamine disease model mice. (2016)
  • Author(s): Kino Y, Washizu C, Kurosawa M, Yamada M, Doi H, Takumi T, Adachi H, Katsuno M, Sobue G, Hicks GG, Hattori N, Shimogori T, Nukina N.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 35236-35236
  • DOI: 10.1038/srep35236
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Differential roles of NF-Y transcription factor in ER chaperone expression and neuronal maintenance in the CNS (2016)
  • Author(s): Yamanaka, T., Tosaki, A., Miyazaki, H., Kurosawa, M., Koike, M., Uchiyama, Y., Maity, SN., Misawa, H., Takahashi, R., Shimogori, T., Hattori, N., Nukina, N.
  • Journal Title: Sci. Rep. Volume: 6 Issue: 1 Pages: 34575-34575
  • DOI: 10.1038/srep34575
  • NAID: 120006349326
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Reappraisal of VAChT-Cre: Preference in slow motor neurons innervating type I or IIa muscle fibers (2016)
  • Author(s): Misawa H, Inomata D, Kikuchi M, Maruyama S, Moriwaki Y, Okuda T, Nukina N, Yamanaka T.
  • Journal Title: Genesis. Volume: 54 Issue: 11 Pages: 568-572
  • DOI: 10.1002/dvg.22979
  • Peer Reviewed
• [Journal Article] Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel β subunits. (2016)
  • Author(s): Shimizu H, Miyazaki H, Ohsawa N, Shoji S, Ishizuka-Katsura Y, Tosaki A, Oyama F, Terada T, Sakamoto K, Shirouzu M, Sekine S, Nukina N, Yokoyama S.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1
  • DOI: 10.1038/srep26618
  • Peer Reviewed / Open Access
• [Journal Article] Acidic mammalian chitinase is a proteases-resistant glycosidase in mouse digestive system. (2016)
  • Author(s): Ohno M, Kimura M, Miyazaki H, Okawa K, Onuki R, Nemoto C, Tabata E, Wakita S, Kashimura A, Sakaguchi M, Sugahara Y, Nukina N, Bauer PO, Oyama F.
  • Journal Title: Sci Rep. Volume: 6
  • Peer Reviewed / Open Access
• [Journal Article] Splicing misregulation of SCN5A contributes to cardiac-conduction delay and heart arrhythmia in myotonic dystrophy. (2016)
  • Author(s): Freyermuth F, Rau F, Kokunai Y, Linke T, Sellier C, Nakamori M, Kino Y,.......Zimmer T, Furling D, Takahashi MP, Charlet-Berguerand N.
  • Journal Title: Nat Commun. Volume: 7 Issue: 1 Pages: 11067-11067
  • DOI: 10.1038/ncomms11067
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Genome-wide analyses in neuronal cells reveal that upstream transcription factors regulate lysosomal gene expression. (2016)
  • Author(s): Yamanaka T, Tosaki A, Kurosawa M, Shimogori T, Hattori N, Nukina N
  • Journal Title: FEBS J Volume: 283(6) Issue: 6 Pages: 1077-87
  • DOI: 10.1111/febs.13650
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] erine 403-phosphorylated p62/SQSTM1 immunoreactivity in inclusions of neurodegenerative diseases (2015)
  • Author(s): Kurosawa,M., Matsumoto,G., Sumikura,H., Hatsuta,H., Murayama,S., Sakurai,T., Shimogori,T., Hattori,N. and Nukina,N.
  • Journal Title: Neurosci. Res. Volume: 103 Pages: 64-70
  • DOI: 10.1016/j.neures.2015.08.002
  • Peer Reviewed / Open Access
• [Journal Article] The Parkinson's Disease-Associated Protein Kinase LRRK2 Modulates Notch Signaling through the Endosomal Pathway. (2015)
  • Author(s): Imai Y, Kobayashi Y, Inoshita T, Meng H, Arano T, Uemura K, Asano T, Yoshimi K, Zhang CL, Matsumoto G, Ohtsuka T, Kageyama R, Kiyonari H, Shioi G, Nukina N, Hattori N, Takahashi R.
  • Journal Title: PLoS Genet. Volume: 11
  • NAID: 120005946994
  • Peer Reviewed / Open Access
• [Journal Article] FUS/TLS deficiency causes behavioral and pathological abnormalities distinct from amyotrophic lateral sclerosis. (2015)
  • Author(s): Kino Y, Washizu C, Kurosawa M, Yamada M, Miyazaki H, Akagi T, Hashikawa T, Doi H, Takumi T, Hicks GG, Hattori N, Shimogori T, Nukina N.
  • Journal Title: Acta Neuropathol Commun. Volume: 3 Pages: 24-24
  • Peer Reviewed / Open Access
• [Journal Article] TBK1 controls autophagosomal engulfment of polyubiquitinated mitochondria through p62/SQSTM1 phosphorylation. (2015)
  • Author(s): Matsumoto G, Shimogori T, Hattori N, Nukina N.
  • Journal Title: Hum Mol Genet. Volume: 24 Issue: 15 Pages: 4429-42
  • DOI: 10.1093/hmg/ddv179
  • Peer Reviewed
• [Presentation] Cell type-specific neruopathologies in NF-Y mutant mice. (2015)
  • Author(s): 山中智行、戸崎麻子、宮崎晴子、黒澤大、小池正人、内山安男、Sankar N. Maity、三澤日出巳、服部信孝、貫名信行
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸（神戸国際会議場）
  • Year and Date: 2015-07-28
• [Remarks] 同志社大学大学院脳科学研究科認知記憶加齢部門
  • URL: http://brainscience.doshisha.ac.jp/introduction/pat/ca.html
• [Remarks] 同志社大学大学院脳科学研究科貫名研究室
  • URL: http://snp.hippy.jp/