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	Cas12a domain flexibility guides R-loop formation and forces RuvC resetting.
ジャーナル・号・ページ	Mol Cell, Vol. 84, Issue 14, Page 2717-22731.e6, Year 2024
掲載日	2024年7月25日
著者	Isabel Strohkendl / Aakash Saha / Catherine Moy / Alexander-Hoi Nguyen / Mohd Ahsan / Rick Russell / Giulia Palermo / David W Taylor /
PubMed 要旨	The specific nature of CRISPR-Cas12a makes it a desirable RNA-guided endonuclease for biotechnology and therapeutic applications. To understand how R-loop formation within the compact Cas12a enables ...The specific nature of CRISPR-Cas12a makes it a desirable RNA-guided endonuclease for biotechnology and therapeutic applications. To understand how R-loop formation within the compact Cas12a enables target recognition and nuclease activation, we used cryo-electron microscopy to capture wild-type Acidaminococcus sp. Cas12a R-loop intermediates and DNA delivery into the RuvC active site. Stages of Cas12a R-loop formation-starting from a 5-bp seed-are marked by distinct REC domain arrangements. Dramatic domain flexibility limits contacts until nearly complete R-loop formation, when the non-target strand is pulled across the RuvC nuclease and coordinated domain docking promotes efficient cleavage. Next, substantial domain movements enable target strand repositioning into the RuvC active site. Between cleavage events, the RuvC lid conformationally resets to occlude the active site, requiring re-activation. These snapshots build a structural model depicting Cas12a DNA targeting that rationalizes observed specificity and highlights mechanistic comparisons to other class 2 effectors.
リンク	Mol Cell / PubMed:38955179 / PubMed Central
手法	EM (単粒子)
解像度	3.3 - 3.8 Å
構造データ	40441 8sfh EMDB-40441, PDB-8sfh: WT CRISPR-Cas12a with a 5bp R-loop 手法: EM (単粒子) / 解像度: 3.4 Å 40442 8sfi EMDB-40442, PDB-8sfi: WT CRISPR-Cas12a with a 8bp R-loop 手法: EM (単粒子) / 解像度: 3.5 Å 40443 8sfj EMDB-40443, PDB-8sfj: WT CRISPR-Cas12a with a 10bp R-loop 手法: EM (単粒子) / 解像度: 3.6 Å 40444 8sfl EMDB-40444, PDB-8sfl: WT CRISPR-Cas12a with a 15bp R-loop 手法: EM (単粒子) / 解像度: 3.3 Å 40445 8sfn EMDB-40445, PDB-8sfn: WT CRISPR-Cas12a with a 16bp R-loop and nontarget strand in the RuvC active site. 手法: EM (単粒子) / 解像度: 3.3 Å 40446 8sfo EMDB-40446, PDB-8sfo: WT CRISPR-Cas12a with a 20bp R-loop and nontarget strand in the RuvC active site. 手法: EM (単粒子) / 解像度: 3.3 Å 40447 8sfp EMDB-40447, PDB-8sfp: WT CRISPR-Cas12a with the target strand in the RuvC active site. 手法: EM (単粒子) / 解像度: 3.8 Å 40448 8sfq EMDB-40448, PDB-8sfq: WT CRISPR-Cas12a post nontarget strand-cleavage with the the RuvC active site exposed. 手法: EM (単粒子) / 解像度: 3.5 Å 40449 8sfr EMDB-40449, PDB-8sfr: WT CRISPR-Cas12a post nontarget strand cleavage. 手法: EM (単粒子) / 解像度: 3.5 Å
化合物	ChemComp-MG: Unknown entry / マグネシウムジカチオン
由来	acidaminococcus sp. bv3l6 (バクテリア) synthetic construct (人工物)
キーワード	DNA BINDING PROTEIN/DNA/RNA / CRISPR / R-loop / endonuclease / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA-RNA complex