PMID:19762288

Citation	Manelyte, L, Guy, CP, Smith, RM, Dillingham, MS, McGlynn, P and Savery, NJ (2009) The unstructured C-terminal extension of UvrD interacts with UvrB, but is dispensable for nucleotide excision repair. DNA Repair (Amst.) 8:1300-10
Abstract	During nucleotide excision repair (NER) in bacteria the UvrC nuclease and the short oligonucleotide that contains the DNA lesion are removed from the post-incision complex by UvrD, a superfamily 1A helicase. Helicases are frequently regulated by interactions with partner proteins, and immunoprecipitation experiments have previously indicated that UvrD interacts with UvrB, a component of the post-incision complex. We examined this interaction using 2-hybrid analysis and surface plasmon resonance spectroscopy, and found that the N-terminal domain and the unstructured region at the C-terminus of UvrD interact with UvrB. We analysed the properties of a truncated UvrD protein that lacked the unstructured C-terminal region and found that it showed a diminished affinity for single-stranded DNA, but retained the ability to displace both UvrC and the lesion-containing oligonucleotide from a post-incision nucleotide excision repair complex. The interaction of the C-terminal region of UvrD with UvrB is therefore not an essential feature of the mechanism by which UvrD disassembles the post-incision complex during NER. In further experiments we showed that PcrA helicase from Bacillus stearothermophilus can also displace UvrC and the excised oligonucleotide from a post-incision NER complex, which supports the idea that PcrA performs a UvrD-like function during NER in gram-positive organisms.
Links	PubMed PMC2997466 Online version:10.1016/j.dnarep.2009.08.005
Keywords	Adenosine Triphosphatases/metabolism; DNA/chemistry; DNA/metabolism; DNA Helicases/chemistry; DNA Helicases/genetics; DNA Helicases/metabolism; DNA Repair; DNA-Binding Proteins/metabolism; Escherichia coli/enzymology; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Models, Molecular; Nucleic Acid Conformation; Protein Binding; Protein Structure, Tertiary; Substrate Specificity
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ECOLI:UVRD		GO:0009411: response to UV	ECO:0000315:		P		Figure five shows that strains with a functional copy of uvrD survive drastically better after exposure to UV light.	complete
ECOLI:UVRD		GO:0003678: DNA helicase activity	ECO:0000315:		F		Figure six illustrates that strains lacking the uvrD gene have helicases with a lower affinity for single stranded DNA	complete

References

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