| Project/Area Number |
23790068
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biological pharmacy
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| Research Institution | Hokkaido University |
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Principal Investigator |
MAITA Hiroshi 北海道大学, 薬学研究科(研究院), 講師 (60431318)
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | 分子生物学 / スプライシング / snRNP / スプリットルシフェラーゼ / スプライソソーム / pre-mRNA splicing / spliceosome / split luciferase |
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| Research Abstract |
The spliceosome is a highly dynamic macromolecular ribonucleoprotein (RNP) machine that catalyzes pre-mRNA splicing by assembling U1, U2, U4, U5, and U6 small nuclear (sn)RNPs. To process large numbers of introns, synthesis and recycling of snRNPs must be maintained within an appropriate range to avoid their shortage, which would cause a severe disease such as Retinitis pigmentosa. However, the mechanism that maintains cellular snRNP levels is unknown. We constructed an expression library of a luciferase fragment fused to core components of snRNPs and used it to isolate PRPF6 and U5-40K that specifically reconstitute luciferase activity in the U5 snRNP complex. We revealed that the reporter detects the effects of small molecules on the levels of the U5 snRNP-reporter protein complex. Our assay will be useful to identify the small molecule that can control the snRNP level in an appropriate range.
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