| Project/Area Number |
12307009
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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 | 一般 |
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| Research Field |
Virology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OHKA Seii The University of Tokyo, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (80313097)
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| Co-Investigator(Kenkyū-buntansha) |
KAMOSHITA Nobuhiko The University of Tokyo, The Institute of Medical Science, Research Associate, 医科学研究所, 助手 (90302603)
KUGE Shusuke The University of Tokyo, Graduate School of Medicine, Lecturer, 大学院・医学系研究科, 講師 (50186376)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥36,600,000 (Direct Cost: ¥33,000,000、Indirect Cost: ¥3,600,000)
Fiscal Year 2001: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000)
Fiscal Year 2000: ¥21,000,000 (Direct Cost: ¥21,000,000)
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| Keywords | Poliovirus / Poliovirus Receptor / Cytopathic Effect / Cytoplasmic Dynein / Basolateral Sorting / ポリオウイルスレセプター / 軸索輸送 / 逆行性モーター蛋白質 / clathrin adaptor complex |
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| Research Abstract |
We discovered that two membrane bound isoforms of poliovirus receptor (PVR), PVRα and PVRδ, showed different localization on the cell membrane of polarized epithelial cells. PVRα displayed basolateral localization although PVRδ showed non-polarized distribution. We demonstrated that basolateral sorting of PVRα was due to tyrosine motif in its cytoplasmic tail. This tyrosine motif interacted with μ1B, a subunit of clathrin adaptor complex AP-1B. It is known that μ1B recruits membrane bound molecules bearing the tyrosine motif to basolateral surface. These evidences suggest that μ1B recruits PVRα to basolateral surface by interacting the tyrosine motif of PVRα.
We found that treatment with anti-PV or anti-PVR antibodies prevented cytopathic effect caused by PV infection in neuroblastoma cells. Under these conditions, host protein synthesis once decreased and then recovered to the normal extent. On the contrary, viral protein synthesis once increased and then decreased. It is known that PV 2A protease relates to cleavage of eIF4G, one of the factors for Cap dependent translation. The amount of 2A correlated with that of cleaved eIF4G in the cells. These results indicate the possibility that expression of cytopthic effect is regulated at the stage of translation.
We proposed a hypothesis that PV is endocytosed from the synapse end of neural cells dependent of PVR, and the vesicles containing PV is retrogradely transported by cytoplasmic dynein to the cell body. It was suggested that cytoplasmic dynein interacts with the cytoplasmic tail of PVR. The motif necessary for the interaction was also revealed.
Replication competent PV vector that expresses BDNF was constructed and injected into the central nervous system of PV sensitive transgenic mice. BDNF and PV antigens were expressed specifically at motor neuron in the spinal cord. This vector replicated stably up to at least the third passage.
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