8BD4

TniQ-capped Tns-ATP-dsDNA complex

8BD4 の概要

• エントリーDOI: 10.2210/pdb8bd4/pdb
• EMDBエントリー: 15974
• 分子名称: TnsC, TniQ (Homology model), DNA (5'-D(P*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*C)-3'), ... (6 entities in total)
• 機能のキーワード: transposition, tniq, tnsc, crispr-cas, tn7-like transposon, dna binding protein
• 由来する生物種: Scytonema hofmannii; 詳細
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 291055.28

構造登録者

Querques, I.,Schmitz, M.,Oberli, S.,Chanez, C.,Jinek, M. (登録日: 2022-10-18, 公開日: 2022-12-28, 最終更新日: 2024-07-24)

主引用文献

Schmitz, M.,Querques, I.,Oberli, S.,Chanez, C.,Jinek, M.
Structural basis for the assembly of the type V CRISPR-associated transposon complex.
Cell, 185:4999-, 2022

PubMed Abstract: CRISPR-Cas systems have been co-opted by Tn7-like transposable elements to direct RNA-guided transposition. Type V-K CRISPR-associated transposons rely on the concerted activities of the pseudonuclease Cas12k, the AAA+ ATPase TnsC, the Zn-finger protein TniQ, and the transposase TnsB. Here we present a cryo-electron microscopic structure of a target DNA-bound Cas12k-transposon recruitment complex comprised of RNA-guided Cas12k, TniQ, a polymeric TnsC filament and, unexpectedly, the ribosomal protein S15. Complex assembly, mediated by a network of interactions involving the guide RNA, TniQ, and S15, results in R-loop completion. TniQ contacts two TnsC protomers at the Cas12k-proximal filament end, likely nucleating its polymerization. Transposition activity assays corroborate our structural findings, implying that S15 is a bona fide component of the type V crRNA-guided transposon machinery. Altogether, our work uncovers key mechanistic aspects underpinning RNA-mediated assembly of CRISPR-associated transposons to guide their development as programmable tools for site-specific insertion of large DNA payloads.

PubMed: 36435179
DOI: 10.1016/j.cell.2022.11.009

実験手法

ELECTRON MICROSCOPY (3.44 Å)