Skip to main content
Journal cover image

Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM.

Publication ,  Journal Article
Riccio, AA; Bouvette, J; Longley, MJ; Krahn, JM; Borgnia, MJ; Copeland, WC
Published in: Methods
September 2022

The mitochondrial replisome replicates the 16.6 kb mitochondria DNA (mtDNA). The proper functioning of this multicomponent protein complex is vital for the integrity of the mitochondrial genome. One of the critical protein components of the mitochondrial replisome is the Twinkle helicase, a member of the Superfamily 4 (SF4) helicases. Decades of research has uncovered common themes among SF4 helicases including self-assembly, ATP-dependent translocation, and formation of protein-protein complexes. Some of the molecular details of these processes are still unknown for the mitochondria SF4 helicase, Twinkle. Here, we describe a protocol for expression, purification, and single-particle cryo-electron microscopy of the Twinkle helicase clinical variant, W315L, which resulted in the first high-resolution structure of Twinkle helicase. The methods described here serve as an adaptable protocol to support future high-resolution studies of Twinkle helicase or other SF4 helicases.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Methods

DOI

EISSN

1095-9130

Publication Date

September 2022

Volume

205

Start / End Page

263 / 270

Location

United States

Related Subject Headings

  • Mitochondrial Proteins
  • Mitochondria
  • DNA, Mitochondrial
  • DNA Replication
  • DNA Helicases
  • Cryoelectron Microscopy
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Riccio, A. A., Bouvette, J., Longley, M. J., Krahn, J. M., Borgnia, M. J., & Copeland, W. C. (2022). Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM. Methods, 205, 263–270. https://doi.org/10.1016/j.ymeth.2022.06.012
Riccio, Amanda A., Jonathan Bouvette, Matthew J. Longley, Juno M. Krahn, Mario J. Borgnia, and William C. Copeland. “Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM.Methods 205 (September 2022): 263–70. https://doi.org/10.1016/j.ymeth.2022.06.012.
Riccio AA, Bouvette J, Longley MJ, Krahn JM, Borgnia MJ, Copeland WC. Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM. Methods. 2022 Sep;205:263–70.
Riccio, Amanda A., et al. “Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM.Methods, vol. 205, Sept. 2022, pp. 263–70. Pubmed, doi:10.1016/j.ymeth.2022.06.012.
Riccio AA, Bouvette J, Longley MJ, Krahn JM, Borgnia MJ, Copeland WC. Method for the structural analysis of Twinkle mitochondrial DNA helicase by cryo-EM. Methods. 2022 Sep;205:263–270.
Journal cover image

Published In

Methods

DOI

EISSN

1095-9130

Publication Date

September 2022

Volume

205

Start / End Page

263 / 270

Location

United States

Related Subject Headings

  • Mitochondrial Proteins
  • Mitochondria
  • DNA, Mitochondrial
  • DNA Replication
  • DNA Helicases
  • Cryoelectron Microscopy
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences