4ZWT

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

4ZWT の概要

• エントリーDOI: 10.2210/pdb4zwt/pdb
• 関連するPDBエントリー: 4ZWQ 4ZWR 4ZWS
• 分子名称: Recombination protein uvsY (1 entity in total)
• 機能のキーワード: recombination, dna binding, homo-heptamer, asymmetry, alpha barrel, viral protein
• 由来する生物種: Enterobacteria phage T4
• タンパク質・核酸の鎖数: 14
• 化学式量合計: 250662.75

構造登録者

Gajewski, S.,White, S.W. (登録日: 2015-05-19, 公開日: 2016-03-02, 最終更新日: 2023-09-27)

主引用文献

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

実験手法

X-RAY DIFFRACTION (4.2 Å)