Beatty L G, Babineau-Clary D, Hogrefe C, Sadowski P D
J Mol Biol. 1986 Apr 20;188(4):529-44. doi: 10.1016/s0022-2836(86)80003-1.
The 2-micron plasmid of the yeast Saccharomyces cerevisiae encodes a site-specific recombinase (FLP) that promotes inversion across a unique site contained in each of the 599-base-pair inverted repeats of the plasmid. We have studied the topological changes generated in supercoiled substrates after exposure to the purified FLP protein in vitro. When a supercoiled substrate bearing two FLP target sequences in inverse orientation is treated with FLP, the products are multiply knotted structures that arise as a result of random entrapment of interdomainal supercoils. Likewise, a supercoiled substrate bearing two target sequences in direct orientation yields multiply interlocked catenanes as the product. Both types of substrate seem to be able to undergo repeated rounds of recombination that result in products of further complexity. The FLP protein also acts as a site-specific topoisomerase during the recombination reaction.
酿酒酵母的2微米质粒编码一种位点特异性重组酶（FLP），该酶可促进质粒599个碱基对反向重复序列中每个序列所含独特位点上的反向排列。我们已经研究了在体外将超螺旋底物暴露于纯化的FLP蛋白后所产生的拓扑变化。当用FLP处理带有两个反向FLP靶序列的超螺旋底物时，产物是多结结构，这是由于结构域间超螺旋的随机捕获而产生的。同样，带有两个正向靶序列的超螺旋底物产生多连环体作为产物。两种类型的底物似乎都能够进行重复的重组循环，从而产生更复杂的产物。在重组反应过程中，FLP蛋白还作为位点特异性拓扑异构酶发挥作用。
