IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v16y2025i1d10.1038_s41467-025-60289-0.html
   My bibliography  Save this article

Autoregulation of the real-time kinetics of the human mitochondrial replicative helicase

Author

Listed:
  • Ismael Plaza-G A

    (IMDEA Nanociencia)

  • María Ortiz-Rodríguez

    (IMDEA Nanociencia)

  • Seth P. Buchanan

    (University of North Florida)

  • Samuel Miguez-Amil

    (Spanish National Cancer Research Centre (CNIO))

  • Kateryna M. Lemishko

    (IMDEA Nanociencia
    Quantemol Ltd)

  • Fernando Moreno-Herrero

    (Centro Nacional de Biotecnología (CSIC))

  • Rafael Fernandez-Leiro

    (Spanish National Cancer Research Centre (CNIO))

  • Grzegorz L. Ciesielski

    (University of North Florida)

  • Borja Ibarra

    (IMDEA Nanociencia
    Unidad Asociada al Centro Nacional de Biotecnología (CSIC))

Abstract

The human mitochondrial helicase Twinkle is essential for mitochondrial DNA (mtDNA) replication and integrity. Using biochemical and single-molecule techniques, we investigated Twinkle’s real-time kinetics, including DNA loading, unwinding, and rewinding, and their regulation by its N-terminal Zinc-binding domain (ZBD), C-terminal tail, and mitochondrial SSB protein (mtSSB). Our results indicate that Twinkle rapidly scans dsDNA to locate the fork, where specific interactions halt diffusion. During unwinding, ZBD-DNA interactions and C-terminal tail control of ATPase activity downregulate kinetics, slowing down the helicase. Binding of mtSSB to DNA likely outcompetes ZBD-DNA interactions, alleviating the downregulatory effects of this domain. Furthermore, we show that ZBD-DNA interactions and ATP binding also regulate rewinding kinetics following helicase stalling. Our findings reveal that ZBD and C-terminal tail play a major role in regulation of Twinkle´s real-time kinetics. Their interplay constitutes an auto-regulatory mechanism that may be relevant for coordinating the mtDNA maintenance activities of the helicase.

Suggested Citation

  • Ismael Plaza-G A & María Ortiz-Rodríguez & Seth P. Buchanan & Samuel Miguez-Amil & Kateryna M. Lemishko & Fernando Moreno-Herrero & Rafael Fernandez-Leiro & Grzegorz L. Ciesielski & Borja Ibarra, 2025. "Autoregulation of the real-time kinetics of the human mitochondrial replicative helicase," Nature Communications, Nature, vol. 16(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-60289-0
    DOI: 10.1038/s41467-025-60289-0
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-025-60289-0
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-025-60289-0?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-60289-0. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.