CRISPR-Csm複合体の構造研究は,同転写干渉のメカニズムを明らかにする
Lilan You1, Jun Ma2, Jiuyu Wang3
1Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 100101 Beijing, China; University of Chinese Academy of Sciences, 100049 Beijing, China.
Cell
|December 4, 2018
まとめ
CRISPR-Cas Csm コンプレックス
科学分野:
- 分子生物学
- 構造生物学
- 生物化学
背景:
- CsmコンプレックスはCRISPR-Casの 干渉システムです
- その機能には,RNA誘導によるRNA分裂,DNA分裂,および循環性オリゴアデニlate (cOA) 合成が含まれています.
- これらの機能の標的RNA依存活性化の構造的基礎は不明である.
研究 の 目的:
- 標的RNAによるCsm複合体の活性化に伴う構造的メカニズムを解明する.
- 標的RNA結合がDNA分裂とCOA生成をどのように引き起こすかを理解する.
主な方法:
- Csm複合体の構造を決定するX線結晶学.
- 構造分析のための冷凍電子顕微鏡 (Cryo-EM).
- 酵素活性を評価するための生化学的測定
主要な成果:
- Csm複合体とcrRNAおよび標的RNA (同種および非同種) の構造を決定した.
- 標的RNAの3'抗タグ領域は,Csm複合体の2つの異なる部位に結合する.
- 同性標的RNAの結合は,Csm1の構成変化によるDNA分裂とcOA合成のアロステリック活性化を誘導する.
結論:
- crRNA媒介のRNA分裂,標的RNA依存のDNA分裂,およびcOA生成に関する構造的洞察
- 関連標的RNAによるCsm複合体のアロステリック活性化のメカニズムが解明された.
- タイプIII-AのCRISPR-Casシステムを理解するための枠組みを提供します.
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