PMID:9049317
| Citation |
Gordienko, I and Rupp, WD (1997) The limited strand-separating activity of the UvrAB protein complex and its role in the recognition of DNA damage. EMBO J. 16:889-95 |
|---|---|
| Abstract |
The recognition by Escherichia coli Uvr nucleotide excision repair proteins of a variety of lesions with diverse chemical structures and the presence of helicase activity in the UvrAB complex which can displace short oligonucleotides annealed to single-stranded DNA led to a model in which this activity moves UvrAB along undamaged DNA to damaged sites where the lesion blocks further translocation and the protein-DNA pre-incision complex is formed. To evaluate this mechanism for damage recognition, we constructed substrates with oligonucleotides of different lengths annealed to single-stranded DNA circles and placed a single 2-(acetylamino)fluorene (AAF) lesion either on the oligonucleotide or on the circle. For the substrates with no lesion, the UvrAB complex effectively displaced a 22-mer but not a 27-mer or longer fragments. The presence of AAF on the oligonucleotide significantly increased the release of the 27-mer but oligomers of 30 or longer were not separated. Placing the lesion on the circular strand did not block the release of the fragments. Instead, the releasing activity of UvrAB was stimulated and also depended on the length of the annealed oligonucleotide. These observations do not agree with the predictions of a damage recognition mechanism that depends on helicase-driven translocation. Most likely, the strand-separating activity of UvrAB is a consequence of local changes occurring during the formation of a DNA-protein pre-incision complex at the damaged site and is not due to translocation of the protein along undamaged DNA to locate a lesion. |
| Links |
PubMed PMC1169689 Online version:10.1093/emboj/16.4.889 |
| Keywords |
2-Acetylaminofluorene; Adenosine Triphosphatases/metabolism; Bacterial Proteins/metabolism; DNA Adducts/chemistry; DNA Damage; DNA Helicases; DNA Repair/physiology; DNA, Circular/chemistry; DNA, Circular/metabolism; DNA, Single-Stranded/chemistry; DNA-Binding Proteins/metabolism; Escherichia coli Proteins; Models, Genetic; Nucleic Acid Conformation; Oligodeoxyribonucleotides |
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