Hashimoto K, Yonesaki T
Department of Botany, Faculty of Science, Kyoto University, Japan.
J Biol Chem. 1991 Mar 15;266(8):4883-8.
Three recombination proteins of bacteriophage T4, uvsX, uvsY, and gene 32 proteins, were examined for the formation of a complex with short single-stranded DNA (ssDNA) molecules containing either 24 or 69 nucleotides. Gel-shift assays revealed that either the uvsX or uvsY protein, when present alone, formed a stable complex only with the 69-mer, while the gene 32 protein bound stably to both ssDNAs. However, a characteristic stable complex formed on the 24-mer when both the uvsX and uvsY proteins were present, and the uvsY protein bound to this DNA in the presence of the gene 32 protein. Isolation of the complexes by centrifugation through a glycerol gradient revealed their protein constituents and showed that the uvsX protein-uvsY protein-24-mer ssDNA complex formed even in the presence of excess gene 32 protein. The possible biological significance of these protein-DNA complexes is discussed.
对噬菌体T4的三种重组蛋白uvsX、uvsY和基因32蛋白进行了检测,以观察它们与含有24个或69个核苷酸的短单链DNA(ssDNA)分子形成复合物的情况。凝胶迁移试验表明,uvsX或uvsY蛋白单独存在时,仅与69聚体形成稳定的复合物,而基因32蛋白则与两种ssDNA都稳定结合。然而,当uvsX和uvsY蛋白都存在时,在24聚体上形成了一种特征性的稳定复合物,并且在基因32蛋白存在的情况下,uvsY蛋白与该DNA结合。通过甘油梯度离心分离复合物,揭示了它们的蛋白质成分,并表明即使在存在过量基因32蛋白的情况下,也会形成uvsX蛋白-uvsY蛋白-24聚体ssDNA复合物。讨论了这些蛋白质-DNA复合物可能的生物学意义。