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Physical and functional interaction between the condensin MukB and the decatenase topoisomerase IV in Escherichia coli

机译:大肠杆菌中凝缩蛋白MukB和脱catenase拓扑异构酶IV之间的物理和功能相互作用

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摘要

Proper geometric and topological organization of DNA is essential for all chromosomal processes. Two classes of proteins play major roles in organizing chromosomes: condensin complexes and type II topoisomerases. In Escherichia coli, MukB, a structural maintenance of chromosome-like component of the bacterial condensin, and topoisomerase IV (Topo IV), a type II topoisomerase that decatenates the newly replicated daughter chromosomes, are both essential for chromosome segregation in rapidly growing cells. However, little is known about the interplay between MukB and Topo IV. Here we demonstrate a physical and functional interaction between MukB and ParC, a subunit of Topo IV, in vitro. The site of MukB interaction was located on the C-terminal domain of ParC and a loss-of-interaction mutant, ParC R705E R729A, was isolated. This variant retained full activity as a topoisomerase when reconstituted with ParE to form Topo IV. We show that MukB stimulates the superhelical DNA relaxation activity of wild-type Topo IV, but not that of Topo IV reconstituted with ParC R705E R729A.
机译:DNA的正确几何和拓扑结构对于所有染色体过程都是必不可少的。两类蛋白质在组织染色体中起主要作用:凝缩蛋白复合物和II型拓扑异构酶。在大肠杆菌中,MukB是细菌凝缩蛋白的染色体样成分的结构维持,拓扑异构酶IV(Topo IV)是用来划分新复制的子染色体的II型拓扑异构酶,对于快速生长的细胞中的染色体分离都是必不可少的。但是,人们对MukB和Topo IV之间的相互作用知之甚少。在这里,我们展示了MukB和ParC(Topo IV的一个亚基)之间的物理和功能相互作用。 MukB相互作用的位点位于ParC的C末端结构域,并分离出相互作用丧失的突变体ParC R705E R729A。当与ParE重组形成Topo IV时,该变体保留了作为拓扑异构酶的全部活性。我们表明,MukB刺激野生型Topo IV的超螺旋DNA松弛活性,但不是用ParC R705E R729A重构的Topo IV的超螺旋DNA松弛活性。

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