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TitleStructural and dynamic basis of DNA capture and translocation by mitochondrial Twinkle helicase.
Journal, issue, pagesNucleic Acids Res, Vol. 50, Issue 20, Page 11965-11978, Year 2022
Publish dateNov 11, 2022
AuthorsZhuo Li / Parminder Kaur / Chen-Yu Lo / Neil Chopra / Jamie Smith / Hong Wang / Yang Gao /
PubMed AbstractTwinkle 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- ...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.
External linksNucleic Acids Res / PubMed:36400570 / PubMed Central
MethodsEM (single particle)
Resolution3.51 - 8.5 Å
Structure data

27842

8e2l

EMDB-27842, PDB-8e2l:
Structure of Lates calcarifer Twinkle helicase with ATP and DNA
Method: EM (single particle) / Resolution: 3.51 Å

EMDB-27843: Structure of Lates calcarifer Twinkle helicase with ATP and DNA, conformer 2
Method: EM (single particle) / Resolution: 7.5 Å

EMDB-27844: Structure of Lates calcarifer Twinkle helicase, apo hexamer
Method: EM (single particle) / Resolution: 7.5 Å

EMDB-27845: Structure of Lates calcarifer Twinkle helicase, apo heptamer
Method: EM (single particle) / Resolution: 8.5 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

Source
  • lates calcarifer (barramundi perch)
  • synthetic construct (others)
KeywordsREPLICATION/DNA / Helicase Mitochondrion / REPLICATION-DNA complex

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