Yonesaki T, Minagawa T
EMBO J. 1985 Dec 1;4(12):3321-7. doi: 10.1002/j.1460-2075.1985.tb04083.x.
Gene uvsX of phage T4 controls genetic recombination and the repair of DNA damage. We have recently purified the gene product, and here describe its properties. The protein has a single-stranded DNA-dependent ATPase activity. It binds efficiently to single- and double-stranded DNAs at 0 degrees C in a cooperative manner. At 30 degree C the double-stranded DNA-protein complex was stable, but the single-stranded DNA-protein complex dissociated rapidly. The instability of the latter complex was reduced by ATP. The protein renatured heat-denatured double-stranded DNA, and assimilated linear single-stranded DNA into homologous superhelical duplexes to produce D-loops. The reaction is stimulated by gene 32 protein when the uvsX protein is limiting. With linear double-stranded DNA and homologous, circular single-stranded DNA, the protein catalyzed single-strand displacement in the 5' to 3' direction with the cooperation of gene 32 protein. All reactions required Mg2+, and all except DNA binding required ATP. We conclude that the uvsX protein is directly involved in strand exchange and is analogous to the recA protein of Escherichia coli. The differences between the uvsX protein and the recA protein, and the role of gene 32 protein in single-strand assimilation and single-strand displacement are briefly discussed.
噬菌体T4的uvsX基因控制着基因重组和DNA损伤修复。我们最近纯化了该基因产物,在此描述其特性。该蛋白质具有单链DNA依赖性ATP酶活性。它在0℃时以协同方式有效结合单链和双链DNA。在30℃时,双链DNA-蛋白质复合物稳定,但单链DNA-蛋白质复合物迅速解离。ATP可降低后一种复合物的不稳定性。该蛋白质能使热变性的双链DNA复性,并将线性单链DNA整合到同源超螺旋双链体中以产生D环。当uvsX蛋白有限时,基因32蛋白可刺激该反应。对于线性双链DNA和同源环状单链DNA,该蛋白质在基因32蛋白的协同作用下催化5'至3'方向的单链置换。所有反应都需要Mg2+,除了DNA结合外,所有反应都需要ATP。我们得出结论,uvsX蛋白直接参与链交换,类似于大肠杆菌的recA蛋白。文中简要讨论了uvsX蛋白与recA蛋白之间的差异,以及基因32蛋白在单链整合和单链置换中的作用。