SPECIFIC BINDING OF THE NIKA PROTEIN TO ONE ARM OF 17-BASE-PAIR INVERTED REPEAT SEQUENCES WITHIN THE ORIT REGION OF PLASMID R64
SPECIFIC BINDING OF THE NIKA PROTEIN TO ONE ARM OF 17-BASE-PAIR INVERTED REPEAT SEQUENCES WITHIN THE ORIT REGION OF PLASMID R64
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DOI：
10.1128/jb.177.1.46-51.1995
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发表时间：
1995-01-01
影响因子：
3.2
通讯作者：
KOMANO, T
中科院分区：
文献类型：
--
作者：
FURUYA, N;KOMANO, T
质粒R64的nikA和nikB基因的产物已显示与R64 oriT DNA形成松弛复合物，并一起作为oriT特异性切口酶发挥作用。我们纯化了nikA基因的蛋白产物。纯化的NikA蛋白与R64 DNA的oriT区域特异性结合。凝胶阻滞试验和DNA酶I足迹分析表明，NikA蛋白只结合到右臂的17 bp的反向重复序列;右臂不同于左臂的一个核苷酸。结合位点邻近切口位点并且在44-bp oriT核心序列内。NikA蛋白的结合诱导R64 oriT序列内的DNA弯曲。
Products of the nikA and nikB genes of plasmid R64 have been shown to form a relaxation complex with R64 oriT DNA and to function together as an oriT-specific nickase. We purified the protein product of the nikA gene. The purified NikA protein bound specifically to the oriT region of R64 DNA. Gel retardation assays and DNase I footprinting analyses indicated that the NikA protein bound only to the right arm of 17-bp inverted repeat sequences; the right arm differed from the left arm by a single nucleotide. The binding site is proximal to the nick site and within the 44-bp oriT core sequence. Binding of the NikA protein induced DNA bending within the R64 oriT sequence.