EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Structural basis of RNA-guided DNA integration by type I CRISPR-associated transposases.
ジャーナル・号・ページ	bioRxiv, Year 2026
掲載日	2026年5月18日
著者	Giada Finocchio / Seraina Oberli / George Lampe / Michael Schmitz / Samuel H Sternberg / Martin Jinek
PubMed 要旨	CRISPR-associated transposases (CASTs) achieve site-specific DNA integration by coupling the RNA-guided targeting action of a nuclease-deficient CRISPR-Cas system with the assembly of a Tn7-like ...CRISPR-associated transposases (CASTs) achieve site-specific DNA integration by coupling the RNA-guided targeting action of a nuclease-deficient CRISPR-Cas system with the assembly of a Tn7-like transpososome complex . Understanding the detailed mechanisms of this elaborate process is paramount to engineering CAST systems into programmable genetic tools . The type I-F CAST ( CAST) displays the highest activity in mammalian cells to date and has been the subject of extensive directed evolution , but efforts to rationally engineer further improvements have been hampered by critical gaps in our understanding of transpososome assembly and activation . Here we use cryo-EM structural analysis, validated by DNA transposition assays, to visualize the CAST system in a series of functional states that define the stepwise mechanism of RNA-guided DNA integration. The structure of a target DNA-bound Cascade-TniQ-TnsC complex reveals that conformational changes induced by R-loop formation are coupled to target DNA stabilization and TnsC heptamerization, which in turn recruits the TnsAB transposase via conserved interactions with its C-terminal tail. Finally, the structure of the 1.2 MDa CAST transpososome holocomplex reveals specific TnsC-TnsB and TnsB-target DNA interactions that drive allosteric remodelling of the TnsB catalytic site to activate donor DNA integration. Together, these findings establish a unified structural and mechanistic blueprint for RNA-guided DNA integration and lay the foundation for engineering next-generation DNA insertion systems for genome editing applications.
リンク	bioRxiv / PubMed:42239233 / PubMed Central
手法	EM (単粒子)
解像度	2.8 - 3.4 Å
構造データ	EMDB-57728: TnsAB-focused cryo-EM volume of the PseCascade-TniQ-TnsC-TnsAB holocomplex 手法: EM (単粒子) / 解像度: 3.4 Å EMDB-57729: TnsC-focused cryo-EM volume of the PseCascade-TniQ-TnsC-TnsAB holocomplex 手法: EM (単粒子) / 解像度: 3.3 Å EMDB-57730: Cascade-TniQ-TnsC-focused cryo-EM volume of the PseCascade-TniQ-TnsC-TnsAB holocomplex 手法: EM (単粒子) / 解像度: 3.1 Å EMDB-57731: Consensus cryo-EM volume of the PseCascade-TniQ-TnsC-TnsAB holocomplex 手法: EM (単粒子) / 解像度: 3.1 Å 57736 30ga EMDB-57736, PDB-30ga: Cryo-EM structure of the PseCascade-TniQ-TnsC-TnsAB holocomplex 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-57737: TnsC-focused cryo-EM volume of the PseCascade-TniQ-TnsC complex bound to TnsB-hook motifs 手法: EM (単粒子) / 解像度: 3.0 Å EMDB-57738: Consensus cryo-EM volume of the PseCascade-TniQ-TnsC complex bound to TnsB-hook motifs 手法: EM (単粒子) / 解像度: 2.8 Å 57739 30gb EMDB-57739, PDB-30gb: Cryo-EM structure of the PseCascade-TniQ-TnsC complex bound to PseTnsB-hook motifs 手法: EM (単粒子) / 解像度: 2.9 Å EMDB-57750: TnsC-focused cryo-EM volume of the PseCascade-TniQ-TnsC complex 手法: EM (単粒子) / 解像度: 3.4 Å EMDB-57751: Consensus cryo-EM volume of the PseCascade-TniQ-TnsC complex 手法: EM (単粒子) / 解像度: 3.1 Å EMDB-57757: Cryo-EM volume of the PseCascade-TniQ complex 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-57758: Cryo-EM volume of the PseCascade complex 手法: EM (単粒子) / 解像度: 2.9 Å 57765 30gt EMDB-57765, PDB-30gt: Cryo-EM structure of the PseCascade-TniQ-TnsC complex 手法: EM (単粒子) / 解像度: 3.3 Å
化合物	ChemComp-MG: Unknown entry ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP, エネルギー貯蔵分子*YM
由来	pseudoalteromonas sp. s983 (バクテリア) escherichia coli k-12 (大腸菌)
キーワード	DNA BINDING PROTEIN / Transposase / DNA-binding / RNA-binding / CRISPR-Cas / CRISPR-associated transposon