8E2L

Structure of Lates calcarifer Twinkle helicase with ATP and DNA

8E2L の概要

• エントリーDOI: 10.2210/pdb8e2l/pdb
• EMDBエントリー: 27842 27843 27844 27845
• 分子名称: Twinkle mtDNA helicase, DNA (5'-D(P*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*T)-3'), ADENOSINE-5'-TRIPHOSPHATE, ... (4 entities in total)
• 機能のキーワード: helicase mitochondrion, replication-dna complex, replication/dna
• 由来する生物種: Lates calcarifer (barramundi perch); 詳細
• タンパク質・核酸の鎖数: 7
• 化学式量合計: 374964.33

構造登録者

Gao, Y.,Li, Z. (登録日: 2022-08-15, 公開日: 2022-11-02, 最終更新日: 2024-06-12)

主引用文献

Li, Z.,Kaur, P.,Lo, C.Y.,Chopra, N.,Smith, J.,Wang, H.,Gao, Y.
Structural and dynamic basis of DNA capture and translocation by mitochondrial Twinkle helicase.
Nucleic Acids Res., 50:11965-11978, 2022

PubMed Abstract: Twinkle is a mitochondrial replicative helicase which can self-load onto and unwind mitochondrial DNA. Nearly 60 mutations on Twinkle have been linked to human mitochondrial diseases. Using cryo-electron microscopy (cryo-EM) and high-speed atomic force microscopy (HS-AFM), we obtained the atomic-resolution structure of a vertebrate Twinkle homolog with DNA and captured in real-time how Twinkle is self-loaded onto DNA. Our data highlight the important role of the non-catalytic N-terminal domain of Twinkle. The N-terminal domain directly contacts the C-terminal helicase domain, and the contact interface is a hotspot for disease-related mutations. Mutations at the interface destabilize Twinkle hexamer and reduce helicase activity. With HS-AFM, we observed that a highly dynamic Twinkle domain, which is likely to be the N-terminal domain, can protrude ∼5 nm to transiently capture nearby DNA and initialize Twinkle loading onto DNA. Moreover, structural analysis and subunit doping experiments suggest that Twinkle hydrolyzes ATP stochastically, which is distinct from related helicases from bacteriophages.

PubMed: 36400570
DOI: 10.1093/nar/gkac1089

実験手法

ELECTRON MICROSCOPY (3.51 Å)