Department of Chemistry, New York University, New York, New York, USA.
Biophys J. 2011 Sep 21;101(6):1403-11. doi: 10.1016/j.bpj.2011.08.011. Epub 2011 Sep 20.
It was found recently that DNA catenanes, formed during replication of circular plasmids, become positively (+) supercoiled, and the unlinking of such catenanes by type IIA topoisomerases proceeds much more efficiently than the unlinking of negatively (-) supercoiled catenanes. In an attempt to explain this striking finding we studied, by computer simulation, conformational properties of supercoiled DNA catenanes. Although the simulation showed that conformational properties of (+) and (-) supercoiled replication catenanes are very different, these properties per se do not give any advantage to (+) supercoiled over (-) supercoiled DNA catenanes for unlinking. An advantage became evident, however, when we took into account the established features of the enzymatic reaction catalyzed by the topoisomerases. The enzymes create a sharp DNA bend in the first bound DNA segment and allow for the transport of the second segment only from inside the bend to its outside. We showed that in (-) supercoiled DNA catenanes this protein-bound bent segment becomes nearly inaccessible for segments of the other linked DNA molecule, inhibiting the unlinking.
最近发现,在环形质粒复制过程中形成的 DNA 连环体变得正超螺旋化,而 IIA 拓扑异构酶对这种连环体的解连环作用比负超螺旋化的连环体的解连环作用效率更高。为了解释这一惊人的发现,我们通过计算机模拟研究了超螺旋化 DNA 连环体的构象特性。尽管模拟表明正超螺旋化和负超螺旋化复制连环体的构象特性非常不同,但这些特性本身并不能为正超螺旋化 DNA 连环体相对于负超螺旋化 DNA 连环体的解连环提供任何优势。然而,当我们考虑到拓扑异构酶催化的酶促反应的既定特征时,优势变得明显。酶在第一个结合的 DNA 片段中创建一个尖锐的 DNA 弯曲,并允许第二个片段仅从弯曲的内部运输到外部。我们表明,在负超螺旋化的 DNA 连环体中,这个与蛋白质结合的弯曲片段几乎无法接近另一个连接的 DNA 分子的片段,从而抑制了解连环。