4ZWR

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

Summary for 4ZWR

• Entry DOI: 10.2210/pdb4zwr/pdb
• Related: 4ZWQ 4ZWS 4ZWT
• Descriptor: Recombination protein uvsY (1 entity in total)
• Functional Keywords: recombination, dna binding, homo-heptamer, asymmetry, alpha barrel, viral protein
• Biological source: Enterobacteria phage T4
• Total number of polymer chains: 7
• Total formula weight: 128554.22

Authors

Gajewski, S.,White, S.W. (deposition date: 2015-05-19, release date: 2016-03-02, Last modification date: 2019-12-04)

Primary citation

Gajewski, S.,Waddell, M.B.,Vaithiyalingam, S.,Nourse, A.,Li, Z.,Woetzel, N.,Alexander, N.,Meiler, J.,White, S.W.
Structure and mechanism of the phage T4 recombination mediator protein UvsY.
Proc.Natl.Acad.Sci.USA, 113:3275-3280, 2016

PubMed Abstract: 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.

PubMed: 26951671
DOI: 10.1073/pnas.1519154113

Experimental method

X-RAY DIFFRACTION (3.3857 Å)