| Budget Amount *help |
¥172,900,000 (Direct Cost: ¥133,000,000、Indirect Cost: ¥39,900,000)
Fiscal Year 2018: ¥34,060,000 (Direct Cost: ¥26,200,000、Indirect Cost: ¥7,860,000)
Fiscal Year 2017: ¥34,060,000 (Direct Cost: ¥26,200,000、Indirect Cost: ¥7,860,000)
Fiscal Year 2016: ¥34,060,000 (Direct Cost: ¥26,200,000、Indirect Cost: ¥7,860,000)
Fiscal Year 2015: ¥33,930,000 (Direct Cost: ¥26,100,000、Indirect Cost: ¥7,830,000)
Fiscal Year 2014: ¥36,790,000 (Direct Cost: ¥28,300,000、Indirect Cost: ¥8,490,000)
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| Outline of Final Research Achievements |
Using the effector complex of RNA interference, called "RISC," as a model system, we developed new techniques that can monitor the molecular dynamics of ncRNA machinery in a real-time and quantitative manner at the single-molecule level. We used these techniques to study the dynamic structural changes of Argonaute proteins, the core of RISC. We found that the structure of Argonaute, which is closed in the empty state, is first opened by helper factors. After incorporating the small RNAs, Argonaute takes a fixed, semi-open structure. Moreover, our data provided molecular insights into how RISC can recognize and cleave their targets so quickly and accurately. In conclusion, single-molecule imaging allowed us to uncover these detailed molecular mechanisms, which were not readily approachable by conventional biochemistry.
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