Abstract
Nucleotide excision repair (NER) is the primary mechanism for removal of ultraviolet light (UV)-induced DNA photoproducts and is mechanistically conserved across all kingdoms of life. Bacterial NER involves damage recognition by UvrA2 and UvrB, followed by UvrC-mediated incision either side of the lesion. Here, using a combination of in vitro and in vivo single-molecule studies we show that a UvrBC complex is capable of lesion identification in the absence of UvrA. Single-molecule analysis of eGFP-labelled UvrB and UvrC in living cells showed that UV damage caused these proteins to switch from cytoplasmic diffusion to stable complexes on DNA. Surprisingly, ectopic expression of UvrC in a uvrA deleted strain increased UV survival. These data provide evidence for a previously unrealized mechanism of survival that can occur through direct lesion recognition by a UvrBC complex.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / deficiency
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Adenosine Triphosphatases / genetics*
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Bacillus / chemistry
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Bacillus / genetics
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Bacillus / metabolism
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DNA Damage
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DNA Helicases / genetics*
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DNA Helicases / metabolism
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DNA Repair*
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DNA, Bacterial / genetics*
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DNA, Bacterial / metabolism
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DNA-Binding Proteins / deficiency
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DNA-Binding Proteins / genetics*
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Endodeoxyribonucleases / genetics*
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Endodeoxyribonucleases / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Escherichia coli / radiation effects*
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Escherichia coli Proteins / genetics*
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Escherichia coli Proteins / metabolism
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Gene Deletion
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Gene Expression Regulation, Bacterial
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Genes, Reporter
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Microbial Viability / genetics
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Microbial Viability / radiation effects
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Protein Binding
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Single Molecule Imaging / methods
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Ultraviolet Rays
Substances
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DNA, Bacterial
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DNA-Binding Proteins
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Escherichia coli Proteins
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UvrB protein, E coli
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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Endodeoxyribonucleases
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UvrC protein, E coli
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UvrA protein, E coli
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Adenosine Triphosphatases
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DNA Helicases