Hargreaves D, Rafferty J B, Sedelnikova S E, Lloyd R G, Rice D W
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, Sheffield S10 2TN, England.
Acta Crystallogr D Biol Crystallogr. 1999 Jan;55(Pt 1):263-5. doi: 10.1107/S0907444998006672. Epub 1999 Jan 1.
During homologous recombination in Escherichia coli the RuvA, B and C proteins interact specifically with the Holliday junction formed by the action of RecA to promote the strand-exchange reaction. RuvA, a homotetrameric protein of molecular weight 88 kDa, has been overexpressed in E. coli, purified and co-crystallized with a synthetic Holliday junction substrate made from four 18-base deoxyoligonucleotides. Crystals were grown using the hanging-drop vapour-diffusion method with sodium acetate as the precipitant. The crystals diffract to a resolution of 6 A and belong to the monoclinic system, space group C2, with cell parameters a = 148, b = 148, c = 106 A and beta = 123 degrees. The X-ray analysis of these crystals should reveal the structure of the Holliday junction and its mode of binding to RuvA, providing new insights into the molecular mechanism of genetic recombination.
在大肠杆菌的同源重组过程中，RuvA、B和C蛋白与RecA作用形成的霍利迪连接体特异性相互作用，以促进链交换反应。RuvA是一种分子量为88 kDa的同四聚体蛋白，已在大肠杆菌中过表达、纯化，并与由四个18碱基的脱氧寡核苷酸制成的合成霍利迪连接体底物共结晶。使用乙酸钠作为沉淀剂，通过悬滴气相扩散法生长晶体。这些晶体的衍射分辨率为6 Å，属于单斜晶系，空间群为C2，晶胞参数a = 148、b = 148、c = 106 Å，β = 123°。对这些晶体的X射线分析应能揭示霍利迪连接体的结构及其与RuvA的结合模式，为遗传重组的分子机制提供新的见解。
