Harris L D, Griffith J D
Lineberger Cancer Research Center, University of North Carolina, Chapel Hill 27514.
J Mol Biol. 1989 Mar 5;206(1):19-27. doi: 10.1016/0022-2836(89)90520-2.
The uvsX and uvsY genes are essential to genetic recombination, recombination-dependent DNA synthesis and to the repair of DNA damage in bacteriophage T4. Purified UvsX protein has been shown to catalyze strand exchange and D-loop formation in vitro, but the role of UvsY protein has been unclear. We report that UvsY protein enhances strand exchange by UvsX protein by interacting specifically with UvsX protein: gene 32 protein (gp32) is not necessary for this effect and UvsY protein has no similar effect on the RecA protein of E. coli. UvsY protein, like UvsX protein, protects single-stranded DNA from digestion by nucleases, but, unlike UvsX protein, shows no ability to protect double-stranded DNA. UvsY protein enhances the rate of single-stranded-DNA-dependent ATP hydrolysis by UvsX protein, particularly in the presence of gp32 or high concentrations of salt, factors that otherwise reduce the ATPase activity of UvsX protein. The enhancement of ATP hydrolysis by UvsY protein is shown to result from the ability of UvsY protein to increase the affinity of UvsX protein for single-stranded DNA.
uvsX和uvsY基因对于噬菌体T4中的基因重组、依赖重组的DNA合成以及DNA损伤修复至关重要。纯化的UvsX蛋白已被证明在体外可催化链交换和D环形成,但UvsY蛋白的作用尚不清楚。我们报告称,UvsY蛋白通过与UvsX蛋白特异性相互作用来增强UvsX蛋白的链交换:基因32蛋白(gp32)对这种效应并非必需,且UvsY蛋白对大肠杆菌的RecA蛋白没有类似作用。与UvsX蛋白一样,UvsY蛋白可保护单链DNA不被核酸酶消化,但与UvsX蛋白不同的是,它没有保护双链DNA的能力。UvsY蛋白可增强UvsX蛋白依赖单链DNA的ATP水解速率,尤其是在存在gp32或高浓度盐的情况下,否则这些因素会降低UvsX蛋白的ATP酶活性。结果表明,UvsY蛋白对ATP水解的增强作用源于其增加UvsX蛋白对单链DNA亲和力的能力。