IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v7y2016i1d10.1038_ncomms12568.html
   My bibliography  Save this article

Single-molecule imaging of UvrA and UvrB recruitment to DNA lesions in living Escherichia coli

Author

Listed:
  • Mathew Stracy

    (University of Oxford
    Biological Physics Research Group, Clarendon Laboratory, University of Oxford)

  • Marcin Jaciuk

    (Laboratory of Protein Structure, International Institute of Molecular and Cell Biology)

  • Stephan Uphoff

    (University of Oxford)

  • Achillefs N. Kapanidis

    (Biological Physics Research Group, Clarendon Laboratory, University of Oxford)

  • Marcin Nowotny

    (Laboratory of Protein Structure, International Institute of Molecular and Cell Biology)

  • David J. Sherratt

    (University of Oxford)

  • Pawel Zawadzki

    (University of Oxford
    Present address: Molecular Biophysics Division, Faculty of Physics, A. Mickiewicz University, Umultowska 85, 61-614 Poznań, Poland)

Abstract

Nucleotide excision repair (NER) removes chemically diverse DNA lesions in all domains of life. In Escherichia coli, UvrA and UvrB initiate NER, although the mechanistic details of how this occurs in vivo remain to be established. Here, we use single-molecule fluorescence imaging to provide a comprehensive characterization of the lesion search, recognition and verification process in living cells. We show that NER initiation involves a two-step mechanism in which UvrA scans the genome and locates DNA damage independently of UvrB. Then UvrA recruits UvrB from solution to the lesion. These steps are coordinated by ATP binding and hydrolysis in the ‘proximal’ and ‘distal’ UvrA ATP-binding sites. We show that initial UvrB-independent damage recognition by UvrA requires ATPase activity in the distal site only. Subsequent UvrB recruitment requires ATP hydrolysis in the proximal site. Finally, UvrA dissociates from the lesion complex, allowing UvrB to orchestrate the downstream NER reactions.

Suggested Citation

  • Mathew Stracy & Marcin Jaciuk & Stephan Uphoff & Achillefs N. Kapanidis & Marcin Nowotny & David J. Sherratt & Pawel Zawadzki, 2016. "Single-molecule imaging of UvrA and UvrB recruitment to DNA lesions in living Escherichia coli," Nature Communications, Nature, vol. 7(1), pages 1-9, November.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms12568
    DOI: 10.1038/ncomms12568
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/ncomms12568
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/ncomms12568?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:7:y:2016:i:1:d:10.1038_ncomms12568. 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.