5DQU

Crystal Structure of Cas-DNA-10 complex

5DQU の概要

• エントリーDOI: 10.2210/pdb5dqu/pdb
• 関連するPDBエントリー: 5DLJ 5DQT 5DQZ
• 分子名称: CRISPR-associated endonuclease Cas1, CRISPR-associated endoribonuclease Cas2, DNA (5'-D(*GP*AP*GP*TP*CP*GP*AP*TP*GP*CP*TP*TP*TP*TP*T)-3'), ... (4 entities in total)
• 機能のキーワード: protein-dna complex, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Escherichia coli K12; 詳細
• 細胞内の位置: Cytoplasm : Q46896
• タンパク質・核酸の鎖数: 10
• 化学式量合計: 170434.81

構造登録者

Wang, J.,Li, J.,Zhao, H.,Sheng, G.,Wang, M.,Yin, M.,Wang, Y. (登録日: 2015-09-15, 公開日: 2015-11-11, 最終更新日: 2023-11-08)

主引用文献

Wang, J.,Li, J.,Zhao, H.,Sheng, G.,Wang, M.,Yin, M.,Wang, Y.
Structural and Mechanistic Basis of PAM-Dependent Spacer Acquisition in CRISPR-Cas Systems.
Cell, 163:840-853, 2015

PubMed Abstract: Bacteria acquire memory of viral invaders by incorporating invasive DNA sequence elements into the host CRISPR locus, generating a new spacer within the CRISPR array. We report on the structures of Cas1-Cas2-dual-forked DNA complexes in an effort toward understanding how the protospacer is sampled prior to insertion into the CRISPR locus. Our study reveals a protospacer DNA comprising a 23-bp duplex bracketed by tyrosine residues, together with anchored flanking 3' overhang segments. The PAM-complementary sequence in the 3' overhang is recognized by the Cas1a catalytic subunits in a base-specific manner, and subsequent cleavage at positions 5 nt from the duplex boundary generates a 33-nt DNA intermediate that is incorporated into the CRISPR array via a cut-and-paste mechanism. Upon protospacer binding, Cas1-Cas2 undergoes a significant conformational change, generating a flat surface conducive to proper protospacer recognition. Here, our study provides important structure-based mechanistic insights into PAM-dependent spacer acquisition.

PubMed: 26478180
DOI: 10.1016/j.cell.2015.10.008

実験手法

X-RAY DIFFRACTION (4.5 Å)