Tse Y, Wang J C
Cell. 1980 Nov;22(1 Pt 1):269-76. doi: 10.1016/0092-8674(80)90174-9.
Escherichia coli and Micrococcus luteus DNA topoisomerase I are found to promote catenation of double-stranded DNA rings. At low DNA concentration dimeric catenanes are the major catenated products; at high DNA concentration or when spermidine is present, catenanes containing more than two rings are formed. There is no requirement of extensive sequence homology between the conponent rings forming a catenane; dimeric catenanes between Pseudomonas phage PM2 DNA and E. coli plasmid pBR322 are readily formed. The formation of a dimeric catenane by these type I topoisomerases, however, requires the presence of at least one preexisting single-chain scission in one of the two component rings. This is in contrast to the cases with the type II DNA topoisomerases which can form catenanes made of covalently closed rings only. The catenanes formed by the type I enzymes can be unlinked by the same enzymes, or by DNA gyrase, a type II enzyme, upon dilution of the isolated catenanes. The catenation and decatenation of duplex DNA rings adds a fourth type of reaction promoted by these type I DNA topoisomerases to the three reported previously: relaxation of superhelical DNA, interconversion between single-stranded DNA rings with and without knots and the intertwining of single-stranded DNA rings of complementary sequences into a covalently closed duplex ring with a high linking number. All four topoisomerization reactions involve the crossing of one DNA strand through a transient break of another DNA strand. The new reaction reported here suggests that such a crossover event might not require pairing of complementary nucleotide sequences.
已发现大肠杆菌和藤黄微球菌DNA拓扑异构酶I可促进双链DNA环的连环化。在低DNA浓度下,二聚连环体是主要的连环化产物;在高DNA浓度下或存在亚精胺时,会形成包含两个以上环的连环体。形成连环体的组成环之间不需要广泛的序列同源性;假单胞菌噬菌体PM2 DNA和大肠杆菌质粒pBR322之间很容易形成二聚连环体。然而,这些I型拓扑异构酶形成二聚连环体需要在两个组成环之一中至少存在一个预先存在的单链断裂。这与II型DNA拓扑异构酶的情况形成对比,后者只能形成由共价闭合环组成的连环体。I型酶形成的连环体在分离的连环体稀释后可被相同的酶或II型酶DNA促旋酶解开。双链DNA环的连环化和解连环化在先前报道的三种反应之外,为这些I型DNA拓扑异构酶促进的反应增加了第四种类型:超螺旋DNA的松弛、有结和无结单链DNA环之间的相互转化以及互补序列的单链DNA环缠绕成具有高连接数的共价闭合双链环。所有四种拓扑异构化反应都涉及一条DNA链穿过另一条DNA链的瞬时断裂。此处报道的新反应表明,这种交叉事件可能不需要互补核苷酸序列配对。