Journal article
Single-molecule imaging of UvrA and UvrB recruitment to DNA lesions in living Escherichia coli
- 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 provide, using single-molecule fluorescence imaging, 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 is dissociated from the lesion complex, allowing UvrB to orchestrate the downstream NER reactions.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 1.2MB, Terms of use)
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- Publisher copy:
- 10.1038/ncomms12568
Authors
+ St John’s College, Oxford
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- Funding agency for:
- Uphoff, S
- Grant:
- Junior Research Fellowship
+ Wellcome Trust
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- Funding agency for:
- Uphoff, S
- Grant:
- Junior Research Fellowship
+ Engineering and Physical Sciences Research Council
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- Funding agency for:
- Stracy, M
- Publisher:
- Nature Publishing Group
- Journal:
- Nature Communications More from this journal
- Volume:
- 7
- Article number:
- 12568
- Publication date:
- 2016-08-26
- Acceptance date:
- 2016-07-14
- DOI:
- ISSN:
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2041-1723
- Pubs id:
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pubs:636302
- UUID:
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uuid:8dd0f729-31fb-4540-9a1b-021d41dc7144
- Local pid:
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pubs:636302
- Source identifiers:
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636302
- Deposit date:
-
2016-07-28
Terms of use
- Copyright holder:
- Stracy et al
- Copyright date:
- 2016
- Notes:
- This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
- Licence:
- CC Attribution (CC BY)
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