| 研究実績の概要 |
We purified histone H1 from nuclear extracts of Drosophila S2 cells. We identified HP1BP3, one of 13 human H1-like chromatin proteins, as a pri-miRNA binding protein that specifically associates with the Microprocessor. Knockdown of HP1BP3, but not other H1 variants, compromised pri-miRNA processing by resulting in the premature release of pri- miRNA transcripts from the chromatin. ChIP studies reveal genome-wide colocalization of HP1BP3 & Drosha and HP1BP3-dependent Drosha binding to actively transcribed miRNA loci. HP1BP3 exhibits a novel pri-miRNA binding activity and promotes the Drosha-pri-miRNA association in vivo. Together, these studies suggest that HP1BP3 promotes co-transcriptional miRNA processing through chromatin retention of nascent pri-miRNA transcripts.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
We have an excellent research team and received help from outstanding collaborators.
1. We collaborated with Dr. Mikiko Siomi at University of Tokyo, who is a world’s leader in small RNA research, to con duct co-localization studies of GFP-TAF11 and Dicer-2/R2D2, using excellent anti-Dicer-2 and anti-R2D2 monoclonal antibo dies that were previously generated in the Siomi lab.
2. Our lab has strong expertise in classical biochemistry. We are good at generating highly purified recombinant Dicer-2 /R2D2 and TAF11 proteins using insect cell-expression system. We developed all of the in vitro assays for analysis of RL C and RISC assembly.
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