9VTP

Target DNA-bound type I-F3 TniQ-Cascade of Vibrio parahaemolyticus in partial R-loop state

9VTP の概要

• エントリーDOI: 10.2210/pdb9vtp/pdb
• EMDBエントリー: 65338
• 分子名称: crRNA from Vibrio parahaemolyticus, Target strand DNA from Vibrio alginolyticus K08M03 plasmid pL300, Non-target strand DNA from Vibrio alginolyticus strain K08M3 plasmid pL300, ... (8 entities in total)
• 機能のキーワード: crispr-associated transposon, dna binding protein
• 由来する生物種: Vibrio parahaemolyticus RIMD 2210633; 詳細
• タンパク質・核酸の鎖数: 13
• 化学式量合計: 491156.67

構造登録者

Ishihara, K.,Numata, T. (登録日: 2025-07-11, 公開日: 2026-02-11)

主引用文献

Ishihara, K.,Matsumoto, S.,Gerle, C.,Gopalasingam, C.C.,Shigematsu, H.,Shirai, T.,Numata, T.
Sequential structural rearrangements at the PAM-distal site of a type I-F3 CRISPR-Cas effector enabling RNA-guided DNA transposition.
Nucleic Acids Res., 54:-, 2026

PubMed Abstract: Some prokaryotes carry CRISPR-associated transposons (CASTs), Tn7-like elements that incorporate genes encoding CRISPR-Cas effectors. CAST insertion is directed by CRISPR-Cas effectors through RNA-guided DNA binding and interactions with transposition-associated proteins. Although efficient sequence-specific DNA integration requires both precise target DNA recognition and coordinated interactions between effectors and transposition-associated proteins, the underlying mechanism remains elusive. Here, we determined three cryo-EM structures of target DNA-bound type I-F3 TniQ-Cascade from Vibrio parahaemolyticus, revealing how Cas8/5 recognizes the protospacer adjacent motif (PAM) and identifying a key residue responsible for the cytidine preference at position -2 of the PAM. We revealed mismatch tolerance at the PAM-proximal site. Structural analyses showed that correct base pairing at the PAM-distal site correlates with conformational changes in the Cas8/5 helical bundle and TniQ, bending the DNA to guide its downstream region toward the transposition machinery. Together, these dynamic rearrangements at the PAM-distal region provide insights into the licensing mechanism of type I-F3 CAST transposition and highlight its potential for genome engineering applications.

PubMed: 41495894
DOI: 10.1093/nar/gkaf1415

実験手法

ELECTRON MICROSCOPY (2.68 Å)