| Project/Area Number |
15K14444
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Molecular biology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
Tadakuma Hisashi 大阪大学, 蛋白質研究所, 助教 (10339707)
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| Research Collaborator |
Kyungmin Baeg
Tsuboyama Kotaro
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | RNAサイレンシング / siRNA / microRNA / 植物 / 自己・非自己認識 / RNA依存性RNAポリメラーゼ |
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| Outline of Final Research Achievements |
RNA-DEPENDENT RNA POLYMERASE 6 (RDR6) is one of the key components for the secondary short interfering RNA (siRNA) biogenesis in plants. Here, we showed that RDR6 specifically converts aberrant non-polyadenylated (non-poly(A)) mRNAs into double-stranded RNAs, which are the precursors of siRNAs. We also demonstrated that the C-terminal RdRP domain of RDR6 possesses a preference for non-poly(A) sequences. Biochemical and biophysical experiments suggest that the poly(A) sequence inhibits the initiation step rather than the elongation step of complementary strand synthesis by RDR6. We have also successfully recapitulated the secondary siRNA biogenesis pathway in the tobacco cell extract. In this system, both the binding of AGO7-RISC to the target mRNA and in vitro translation of SILENCING DEFECTIVE 5 were required for the efficient complementary strand synthesis by RDR6.
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