Nucleotide excision repair (NER) is a major DNA repair mechanism that recognizes a broad range of DNA damages. In Escherichia coli, damage recognition in NER is accomplished by the UvrA and UvrB proteins. We have analysed the structural properties of the different protein-DNA complexes formed by UvrA, UvrB and (damaged) DNA using atomic force microscopy. Analysis of the UvrA(2)B complex in search of damage revealed the DNA to be wrapped around the UvrB protein, comprising a region of about seven helical turns. In the UvrB-DNA pre-incision complex the DNA is wrapped in a similar way and this DNA configuration is dependent on ATP binding. Based on these results, a role for DNA wrapping in damage recognition is proposed. Evidence is presented that DNA wrapping in the pre-incision complex also stimulates the rate of incision by UvrC.

*DNA Repair, *Endodeoxyribonucleases, *Escherichia coli Proteins, Adenosine Triphosphate/metabolism, Adenosinetriphosphatase/chemistry/metabolism, Bacterial Proteins/chemistry/metabolism, Base Sequence, DNA Helicases/chemistry/*metabolism, DNA Primers/genetics, DNA, Bacterial/*chemistry/genetics/*metabolism, DNA-Binding Proteins/chemistry/metabolism, Escherichia coli/genetics/metabolism, Kinetics, Macromolecular Substances, Microscopy, Atomic Force, Molecular Sequence Data, Research Support, Non-U.S. Gov't
dx.doi.org/10.1093/emboj/20.3.601, hdl.handle.net/1765/12916
EMBO Journal
Erasmus MC: University Medical Center Rotterdam

Verhoeven, E.E, Wyman, C, Moolenaar, G.F, Hoeijmakers, J.H.J, & Goosen, N. (2001). Architecture of nucleotide excision repair complexes: DNA is wrapped by UvrB before and after damage recognition. EMBO Journal, 20(3), 601–611. doi:10.1093/emboj/20.3.601