Valens Michèle, Thiel Axel, Boccard Frédéric
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France.
PLoS Genet. 2016 Sep 14;12(9):e1006309. doi: 10.1371/journal.pgen.1006309. eCollection 2016 Sep.
The Ori region of bacterial genomes is segregated early in the replication cycle of bacterial chromosomes. Consequently, Ori region positioning plays a pivotal role in chromosome dynamics. The Ori region of the E. coli chromosome is organized as a macrodomain with specific properties concerning DNA mobility, segregation of loci and long distance DNA interactions. Here, by using strains with chromosome rearrangements and DNA mobility as a read-out, we have identified the MaoP/maoS system responsible for constraining DNA mobility in the Ori region and limiting long distance DNA interactions with other regions of the chromosome. MaoP belongs to a group of proteins conserved in the Enterobacteria that coevolved with Dam methylase including SeqA, MukBEF and MatP that are all involved in the control of chromosome conformation and segregation. Analysis of DNA rings excised from the chromosome demonstrated that the single maoS site is required in cis on the chromosome to exert its effect while MaoP can act both in cis and in trans. The position of markers in the Ori region was affected by inactivating maoP. However, the MaoP/maoS system was not sufficient for positioning the Ori region at the ¼-¾ regions of the cell. We also demonstrate that the replication and the resulting expansion of bulk DNA are localized centrally in the cell. Implications of these results for chromosome positioning and segregation in E. coli are discussed.
细菌基因组的Ori区域在细菌染色体复制周期的早期就被分离出来。因此,Ori区域的定位在染色体动态变化中起着关键作用。大肠杆菌染色体的Ori区域被组织成一个宏观结构域,具有关于DNA移动性、基因座分离和长距离DNA相互作用的特定特性。在这里,通过使用具有染色体重排和DNA移动性作为读出信号的菌株,我们鉴定出了负责限制Ori区域DNA移动性并限制与染色体其他区域长距离DNA相互作用的MaoP/maoS系统。MaoP属于在肠杆菌中保守的一组蛋白质,它们与Dam甲基化酶共同进化,包括SeqA、MukBEF和MatP,这些都参与染色体构象和分离的控制。对从染色体上切除的DNA环的分析表明,单个maoS位点在染色体上顺式发挥作用,而MaoP可以顺式和反式发挥作用。Ori区域中标记的位置受maoP失活的影响。然而,MaoP/maoS系统不足以将Ori区域定位在细胞的1/4 - 3/4区域。我们还证明,大量DNA的复制及其产生的扩展集中在细胞中央。讨论了这些结果对大肠杆菌染色体定位和分离的影响。