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Real-time single-molecule imaging reveals a direct interaction between UvrC and UvrB on DNA tightropes

Hughes, Craig D, Wang, Hong, Ghodke, Harshad, Simons, Michelle, Towheed, Atif, Peng, Ye, Van Houten, Bennett, Kad, Neil M (2013) Real-time single-molecule imaging reveals a direct interaction between UvrC and UvrB on DNA tightropes. Nucleic acids research, 41 (9). pp. 4901-4912. ISSN 0305-1048. E-ISSN 1362-4962. (doi:10.1093/nar/gkt177) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1093/nar/gkt177

Abstract

Nucleotide excision DNA repair is mechanistically conserved across all kingdoms of life. In prokaryotes, this multi-enzyme process requires six proteins: UvrA-D, DNA polymerase I and DNA ligase. To examine how UvrC locates the UvrB-DNA pre-incision complex at a site of damage, we have labeled UvrB and UvrC with different colored quantum dots and quantitatively observed their interactions with DNA tightropes under a variety of solution conditions using oblique angle fluorescence imaging. Alone, UvrC predominantly interacts statically with DNA at low salt. Surprisingly, however, UvrC and UvrB together in solution bind to form the previously unseen UvrBC complex on duplex DNA. This UvrBC complex is highly motile and engages in unbiased one-dimensional diffusion. To test whether UvrB makes direct contact with the DNA in the UvrBC-DNA complex, we investigated three UvrB mutants: Y96A, a ?-hairpin deletion and D338N. These mutants affected the motile properties of the UvrBC complex, indicating that UvrB is in intimate contact with the DNA when bound to UvrC. Given the in vivo excess of UvrB and the abundance of UvrBC in our experiments, this newly identified complex is likely to be the predominant form of UvrC in the cell.

Item Type: Article
DOI/Identification number: 10.1093/nar/gkt177
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: N. Kad
Date Deposited: 15 Sep 2014 19:17 UTC
Last Modified: 29 May 2019 13:05 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/42942 (The current URI for this page, for reference purposes)
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