4zws

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Crystal Structure of the Bacteriophage T4 recombination mediator protein UvsY, Lattice Type IIICrystal Structure of the Bacteriophage T4 recombination mediator protein UvsY, Lattice Type III

Structural highlights

4zws is a 7 chain structure with sequence from Escherichia virus T4. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 2.6Å
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

UVSY_BPT4 Plays a role in viral DNA synthesis by promoting enzymatic activities of UvsX recombinase, by promoting UvsX-ssDNA filament assembly, and by helping UvsX to displace bound gp32 from ssDNA.[1] [2]

Publication Abstract from PubMed

The UvsY recombination mediator protein is critical for efficient homologous recombination in bacteriophage T4 and is the functional analog of the eukaryotic Rad52 protein. During T4 homologous recombination, the UvsX recombinase has to compete with the prebound gp32 single-stranded binding protein for DNA-binding sites and UvsY stimulates this filament nucleation event. We report here the crystal structure of UvsY in four similar open-barrel heptameric assemblies and provide structural and biophysical insights into its function. The UvsY heptamer was confirmed in solution by centrifugation and light scattering, and thermodynamic analyses revealed that the UvsY-ssDNA interaction occurs within the assembly via two distinct binding modes. Using surface plasmon resonance, we also examined the binding of UvsY to both ssDNA and the ssDNA-gp32 complex. These analyses confirmed that ssDNA can bind UvsY and gp32 independently and also as a ternary complex. They also showed that residues located on the rim of the heptamer are required for optimal binding to ssDNA, thus identifying the putative ssDNA-binding surface. We propose a model in which UvsY promotes a helical ssDNA conformation that disfavors the binding of gp32 and initiates the assembly of the ssDNA-UvsX filament.

Structure and mechanism of the phage T4 recombination mediator protein UvsY.,Gajewski S, Waddell MB, Vaithiyalingam S, Nourse A, Li Z, Woetzel N, Alexander N, Meiler J, White SW Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):3275-80. doi:, 10.1073/pnas.1519154113. Epub 2016 Mar 7. PMID:26951671[3]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

  1. Pant K, Shokri L, Karpel RL, Morrical SW, Williams MC. Modulation of T4 gene 32 protein DNA binding activity by the recombination mediator protein UvsY. J Mol Biol. 2008 Jul 25;380(5):799-811. doi: 10.1016/j.jmb.2008.05.039. Epub 2008, May 24. PMID:18565541 doi:http://dx.doi.org/10.1016/j.jmb.2008.05.039
  2. Xu H, Beernink HT, Morrical SW. DNA-binding properties of T4 UvsY recombination mediator protein: polynucleotide wrapping promotes high-affinity binding to single-stranded DNA. Nucleic Acids Res. 2010 Aug;38(14):4821-33. doi: 10.1093/nar/gkq219. Epub 2010, Apr 5. PMID:20371513 doi:http://dx.doi.org/10.1093/nar/gkq219
  3. Gajewski S, Waddell MB, Vaithiyalingam S, Nourse A, Li Z, Woetzel N, Alexander N, Meiler J, White SW. Structure and mechanism of the phage T4 recombination mediator protein UvsY. Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):3275-80. doi:, 10.1073/pnas.1519154113. Epub 2016 Mar 7. PMID:26951671 doi:http://dx.doi.org/10.1073/pnas.1519154113

4zws, resolution 2.60Å

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