| Project/Area Number |
15K06982
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Musashino University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
坂本 泰一 千葉工業大学, 先進工学部, 教授 (40383369)
吉村 明 東北医科薬科大学, 薬学部, 講師 (70302164)
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| Project Period (FY) |
2015-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | スプライシング / がん / pre-mRNAスプライシング / NMR / SELEX / mRNAスプライシング |
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| Outline of Final Research Achievements |
Mutations in some specific lysine residues in a spliceosomal protein SF3b1 have often been found in several types of cancer cells. However, it remains unknown not only what part SF3b1 plays in the splicing reaction, but also how the mutations in SF3b affect the process of splicing reaction and contribute to malignant transformation. To elucidate the function of SF3b1 in the process by structural analysis, we attempted to prepare the recombinant SF3b1 protein by overproduction in E. coli, but the yield of the protein was very low. Next, we examined the interactions between spliceosomal proteins by NMR titration experiments, showing that a spliceosomal protein in which some mutations had been reported in cancer cells makes interactions with another spliceosomal protein. Thus, we tried to determine the core structure of the complex composed of the two proteins by NMR. Furthermore, we performed SELEX to elucidate the specificity of the spliceosomal protein that binds to the branch point.
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| Academic Significance and Societal Importance of the Research Achievements |
本実験では，SF3b1と共に機能し，がん細胞での変異が報告されている特定のスプライシング因子が別のスプライシング因子と相互作用することが判明したため，これらの複合体形成部位の立体構造解析を進めている。これらの相互作用メカニズムの解明は，がん化とスプライシング因子の変異との関係を明らかにするのに非常に役立つと考えられる。また，ブランチ部位に結合するスプライシング因子について，RNA配列の特異性の解明を試みており，ヒトにおける保存性の低いブランチ部位配列を認識するメカニズムに関して，新たな知見が得られると考える。
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