The unstructured C-terminal extension of UvrD interacts with UvrB, but is dispensable for nucleotide excision repair

Laura Manelyte, Colin P Guy, Rachel M Smith, Mark S Dillingham, Peter McGlynn, Nigel J Savery

Research output: Contribution to journalArticle (Academic Journal)peer-review

50 Citations (Scopus)

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.
Translated title of the contributionThe unstructured C-terminal extension of UvrD interacts with UvrB, but is dispensable for nucleotide excision repair
Original languageEnglish
Pages (from-to)1300 - 1310
Number of pages11
JournalDNA Repair
Volume8
Issue number11
DOIs
Publication statusPublished - Nov 2009

Bibliographical note

Other: First published on-line 16th September 2009

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