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)

36 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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