Charman, Robert, Kad, Neil M. (2022) Single molecule iSCAT imaging reveals a fast, energy efficient search mode for the DNA repair protein UvrA. Nanoscale, 14 . pp. 5174-5184. ISSN 2040-3364. E-ISSN 2040-3372. (doi:10.1039/D1NR06913F) (KAR id:93135)
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Official URL: https://doi.org/10.1039/D1NR06913F |
Abstract
Exposure to UV radiation results in numerous DNA lesions, which threaten genome integrity. The nucleotide excision DNA repair pathway detects and repairs a range of such UV-induced DNA lesions. In bacteria, initial damage detection and verification is carried out by two proteins: UvrA and UvrB. Despite decades of study, the process of how these proteins locate damage remains unclear. Here we use high-speed interferometric scattering (iSCAT) microscopy, in combination with a surface-boundDNA assay, to investigate early damage detection by UvrA. We have discovered that UvrA interacts with DNA in two phases; a slow phase (~1.3/s) that correlates with an ATP-consuming state previously identified, and a second, much faster search mode. These faster interactions persist for ~130 ms and using ATP analogues we determine this phase does not require ATP consumption. Including this new fast-search state in a model of the DNA search process reveals that only with this state is it possible for basal levels of UvrA to explore 99% of the E. coli genome within a single division cycle. Altogether, this work uncovers the presence of a rapid, energy efficient search mechanism, which allows UvrA alone to search the entirety of the E. coli genome within a single division cycle.
Item Type: | Article |
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DOI/Identification number: | 10.1039/D1NR06913F |
Subjects: | Q Science |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | Neil Kad |
Date Deposited: | 10 Feb 2022 15:25 UTC |
Last Modified: | 11 Jan 2024 09:25 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/93135 (The current URI for this page, for reference purposes) |
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