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The phage T4 protein UvsW drives Holliday junction branch migration.

Publication ,  Journal Article
Webb, MR; Plank, JL; Long, DT; Hsieh, T-S; Kreuzer, KN
Published in: J Biol Chem
November 23, 2007

The phage T4 UvsW protein has been shown to play a crucial role in the switch from origin-dependent to recombination-dependent replication in T4 infections through the unwinding of origin R-loop initiation intermediates. UvsW also functions with UvsX and UvsY to repair damaged DNA through homologous recombination, and, based on genetic evidence, has been proposed to act as a Holliday junction branch migration enzyme. Here we report the purification and characterization of UvsW. Using oligonucleotide-based substrates, we confirm that UvsW unwinds branched DNA substrates, including X and Y structures, but shows little activity in unwinding linear duplex substrates with blunt or single-strand ends. Using a novel Holliday junction-containing substrate, we also demonstrate that UvsW promotes the branch migration of Holliday junctions efficiently through more than 1000 bp of DNA. The ATP hydrolysis-deficient mutant protein, UvsW-K141R, is unable to promote Holliday junction branch migration. However, both UvsW and UvsW-K141R are capable of stabilizing Holliday junctions against spontaneous branch migration when ATP is not present. Using two-dimensional agarose gel electrophoresis we also show that UvsW acts on T4-generated replication intermediates, including Holliday junction-containing X-shaped intermediates and replication fork-shaped intermediates. Taken together, these results strongly support a role for UvsW in the branch migration of Holliday junctions that form during T4 recombination, replication, and repair.

Duke Scholars

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

November 23, 2007

Volume

282

Issue

47

Start / End Page

34401 / 34411

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Recombination, Genetic
  • Mutation, Missense
  • Membrane Proteins
  • DNA-Binding Proteins
  • DNA, Single-Stranded
  • DNA, Cruciform
  • DNA Replication
  • DNA Repair
 

Citation

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MLA
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Webb, M. R., Plank, J. L., Long, D. T., Hsieh, T.-S., & Kreuzer, K. N. (2007). The phage T4 protein UvsW drives Holliday junction branch migration. J Biol Chem, 282(47), 34401–34411. https://doi.org/10.1074/jbc.M705913200
Webb, Michael R., Jody L. Plank, David T. Long, Tao-shih Hsieh, and Kenneth N. Kreuzer. “The phage T4 protein UvsW drives Holliday junction branch migration.J Biol Chem 282, no. 47 (November 23, 2007): 34401–11. https://doi.org/10.1074/jbc.M705913200.
Webb MR, Plank JL, Long DT, Hsieh T-S, Kreuzer KN. The phage T4 protein UvsW drives Holliday junction branch migration. J Biol Chem. 2007 Nov 23;282(47):34401–11.
Webb, Michael R., et al. “The phage T4 protein UvsW drives Holliday junction branch migration.J Biol Chem, vol. 282, no. 47, Nov. 2007, pp. 34401–11. Pubmed, doi:10.1074/jbc.M705913200.
Webb MR, Plank JL, Long DT, Hsieh T-S, Kreuzer KN. The phage T4 protein UvsW drives Holliday junction branch migration. J Biol Chem. 2007 Nov 23;282(47):34401–34411.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

November 23, 2007

Volume

282

Issue

47

Start / End Page

34401 / 34411

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Recombination, Genetic
  • Mutation, Missense
  • Membrane Proteins
  • DNA-Binding Proteins
  • DNA, Single-Stranded
  • DNA, Cruciform
  • DNA Replication
  • DNA Repair