| Project/Area Number |
16K14696
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Functional biochemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
TASHIMA YUKO 名古屋大学, 医学系研究科, 助教 (10423104)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | 糖脂質GPI / GPIアンカー型タンパク質 / 細胞内輸送 / ゴルジ装置 / 糖鎖 / 輸送 |
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| Outline of Final Research Achievements |
Glycosylphosphatidylinositol (GPI) tether a hydrophilic protein on the plasma membrane. Synthesis of GPI-anchored proteins (GPI-APs) in the ER and its lipid remodeling in the Golgi have been studied very well, but the transport process to the cell surface from the Golgi have not been known well. We newly established a method to monitor the transport of GPI-APs to the cell surface from the Golgi using retention using selective hooks system [Boncompain G et al., Nature methods, 2012] and HEK293 cells. To identify factor proteins for transport of GPI-APs to the cell surface from the Golgi, we used this method by introducing reporter cells with CRISPR/Cas9 and single guide RNA library. We sorted mutant cells that delayed transport of a reporter GPI to the cell surface from the Golgi. The candidate factor proteins to be related with transport of GPI-APs are analyzing by next generation sequencing.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究は、これまで優れたシステムのなかったGPI-APsのゴルジ装置から細胞膜への輸送にフォーカスして、経時的・定量的に解析する系の樹立を試みたことに特色がある。更に、この領域の輸送に働く因子を網羅的に同定することは、GPI-APsの輸送のみならず、その輸送体と考えられているラフトの動態メカニズムの理解にもつながる可能性があり、ラフトの重要性を考慮すると、本研究成果の意義は大きい。また、GPI-APsは、細胞表面に発現した後も、細胞内のエンドソームや細胞外分泌性小胞エクソソームに移動する。これらの輸送のモニターや定量の方法を将来的に開発する土台として、本研究は波及効果が大きい。
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