Crystallographic and NMR analyses of UvsW and UvsW.1 from bacteriophage T4

J Biol Chem. 2007 Nov 23;282(47):34392-400. doi: 10.1074/jbc.M705900200. Epub 2007 Sep 17.

Abstract

The uvsWXY system is implicated in the replication and repair of the bacteriophage T4 genome. Whereas the roles of the recombinase (UvsX) and the recombination mediator protein (UvsY) are known, the precise role of UvsW is unclear. Sequence analysis identifies UvsW as a member of the monomeric SF2 helicase superfamily that translocates nucleic acid substrates via the action of two RecA-like motor domains. Functional homologies to Escherichia coli RecG and biochemical analyses have shown that UvsW interacts with branched nucleic acid substrates, suggesting roles in recombination and the rescue of stalled replication forks. A sequencing error at the 3'-end of the uvsW gene has revealed a second, short open reading frame that encodes a protein of unknown function called UvsW.1. We have determined the crystal structure of UvsW to 2.7A and the NMR solution structure of UvsW.1. UvsW has a four-domain architecture with structural homology to the eukaryotic SF2 helicase, Rad54. A model of the UvsW-ssDNA complex identifies structural elements and conserved residues that may interact with nucleic acid substrates. The NMR solution structure of UvsW.1 reveals a dynamic four-helix bundle with homology to the structure-specific nucleic acid binding module of RecQ helicases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteriophage T4 / enzymology*
  • Crystallography, X-Ray
  • DNA Helicases / chemistry*
  • DNA Helicases / metabolism
  • DNA Repair / physiology
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism
  • Eukaryotic Cells / enzymology
  • Genome, Viral / physiology
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Structure, Secondary / physiology
  • Protein Structure, Tertiary / physiology
  • Rec A Recombinases / chemistry
  • Rec A Recombinases / metabolism
  • RecQ Helicases / chemistry
  • RecQ Helicases / metabolism
  • Recombination, Genetic / physiology
  • Structural Homology, Protein
  • Viral Proteins / chemistry*
  • Viral Proteins / metabolism
  • Virus Replication / physiology

Substances

  • DNA, Single-Stranded
  • Escherichia coli Proteins
  • Viral Proteins
  • RecG protein, E coli
  • Rec A Recombinases
  • DNA Helicases
  • RecQ Helicases
  • UvsW protein, Bacteriophage T4

Associated data

  • PDB/1RIF
  • PDB/2JPN