| Project/Area Number |
08558079
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Cell biology
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| Research Institution | Osaka University |
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Principal Investigator |
YONEDA Yoshihiro Osaka University Medical School, Professor, 医学部, 教授 (80191667)
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| Co-Investigator(Kenkyū-buntansha) |
YUBA Syunsuke Osaka University Medical School, Assistant Professor, 医学部, 助手 (40263248)
HIRAOKA Yasushi Structural Biology Section Kansai Advanced Research Center Communications Research Laboratory, Section Chief, 通信総合研究所・関西先端研究センター・生物情報研究室, 室長
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥18,200,000 (Direct Cost: ¥18,200,000)
Fiscal Year 1998: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1997: ¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1996: ¥9,400,000 (Direct Cost: ¥9,400,000)
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| Keywords | Molecular visualization technique / Nuclear protein transport / Green fluorescent protein / Importin / Fluorescent dye / Nuclear localization signal / Cell cycle / Microinjection / 核-細胞質間物質輸送 / importin / 三次元可視化技術 / 細胞核 / green fluorescent protein / 画像解析 / 三次元可視化 / 光学顕微鏡技術 / 核蛋白質 / 核膜 / 核局在化シグナル |
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| Research Abstract |
We have developed an in vivo assay system which enables us to monitor the behavior of functional molecules three-dimensionally on real time in living cells. Moreover, we have been able to visualize four kinds of molecules such as chromosomeaml proteins and microtubules simultaneously in one living cell. By using this system, we analyzed the behavior of nuclear proteins and nuclear transport factors such as importin α and importin β. As a result, we have obtained the following data.
2. It was found that after mitosis, there exists a short period when the nuclear protein import activity is not yet recovered, even though the integrity of nuclear envelope appears to be restored.
3. When green fluorescent protein (GFP)-tagged importin α was microinjected into the cytoplasm of living cells and the intracellular localization of the GFP-importin α was observed in a live state without fixation, the protein was primarily localized in the cytoplasm. In contrast, when nuclear proteins were injected to the same cells after injection of the GFP-importin α, it was transported into the nucleus within one minute and then returned to the cytoplasm within the next one minute. Moreover, it was found that the overall distribution of the GFP-importin α was altered to the nucleus about two minutes just before mitosis.
4. We found that GFP-importin β was localized throughout the cytoplasm and nucleus and that the overall localization did not change during interphase. Moreover, the injection of nuclear proteins did not affect the intracellular distribution of the GFP-importin β.
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