Morrical S W, Alberts B M
Department of Biochemistry and Biophysics, University of California, San Francisco 94143.
J Biol Chem. 1990 Sep 5;265(25):15096-103.
An in vitro system containing the T4 gene 43, 45, 44/62, 32, dda, and uvsX proteins catalyzes DNA synthesis that is dependent on the synapsis step of homologous genetic recombination. The rate of DNA synthesis in this system is highly dependent on the concentration of the uvsX recombinase (a recA-like protein). Here we report the effect of the T4 uvsY protein, a recombination accessory protein, on this reaction. Low concentrations of uvsY protein greatly stimulate DNA synthesis at low concentrations of uvsX protein, but these same concentrations inhibit DNA synthesis at high concentrations of uvsX protein. As a result, the addition of small amounts of uvsY protein lowers the minimum concentration of uvsX protein needed for the reaction 8-fold, and it lowers the uvsX protein concentration for maximum activity 4-fold. The uvsY protein can affect either the initiation or elongation phase of DNA synthesis, depending on the concentration of uvsX protein present. The implications of these results for the function of the uvsY protein in T4 DNA replication in vivo are discussed.
一个包含T4基因43、45、44/62、32、dda和uvsX蛋白的体外系统催化依赖于同源基因重组联会步骤的DNA合成。该系统中DNA合成的速率高度依赖于uvsX重组酶(一种类RecA蛋白)的浓度。在此,我们报告了重组辅助蛋白T4 uvsY蛋白对该反应的影响。低浓度的uvsY蛋白在低浓度uvsX蛋白时极大地刺激DNA合成,但相同浓度在高浓度uvsX蛋白时却抑制DNA合成。因此,添加少量uvsY蛋白可使反应所需的uvsX蛋白最低浓度降低8倍,并使最大活性时的uvsX蛋白浓度降低4倍。uvsY蛋白可根据存在的uvsX蛋白浓度影响DNA合成的起始或延伸阶段。讨论了这些结果对uvsY蛋白在体内T4 DNA复制功能的意义。