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细菌中姐妹染色单体相互作用的位点特异性重组分析。

Sister chromatid interactions in bacteria revealed by a site-specific recombination assay.

机构信息

Centre de Génétique Moléculaire du CNRS, Gif-sur-Yvette, France.

出版信息

EMBO J. 2012 Aug 15;31(16):3468-79. doi: 10.1038/emboj.2012.194. Epub 2012 Jul 20.

Abstract

The process of Sister Chromosome Cohesion (SCC), which holds together sister chromatids upon replication, is essential for chromosome segregation and DNA repair in eukaryotic cells. Although cohesion at the molecular level has never been described in E. coli, previous studies have reported that sister sequences remain co-localized for a period after their replication. Here, we have developed a new genetic recombination assay that probes the ability of newly replicated chromosome loci to interact physically. We show that Sister Chromatid Interaction (SCI) occurs exclusively within a limited time frame after replication. Importantly, we could differentiate sister cohesion and co-localization since factors such as MatP and MukB that reduced the co-localization of markers had no effect on molecular cohesion. The frequency of sister chromatid interactions were modulated by the activity of Topo-IV, revealing that DNA topology modulates cohesion at the molecular scale in bacteria.

摘要

姐妹染色单体黏合(Sister Chromosome Cohesion,SCC)过程在复制后将姐妹染色单体保持在一起,对于真核细胞中的染色体分离和 DNA 修复至关重要。尽管在大肠杆菌中从未描述过分子水平上的黏合,但先前的研究报告称,姐妹序列在复制后仍会在一段时间内保持共定位。在这里,我们开发了一种新的遗传重组测定法,该方法可探测新复制的染色体位点相互物理作用的能力。我们表明,姐妹染色单体相互作用(Sister Chromatid Interaction,SCI)仅在复制后有限的时间内发生。重要的是,我们可以区分姐妹黏合和共定位,因为诸如 MatP 和 MukB 等降低标记共定位的因素对分子黏合没有影响。拓扑异构酶 IV 的活性调节着姐妹染色单体相互作用的频率,这表明 DNA 拓扑结构在细菌中调节着分子尺度上的黏合。

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