Espeli Olivier, Mercier Romain, Boccard Frédéric
Centre de Génétique Moléculaire du CNRS, Avenue de la Terrasse, 91198 Gif-sur-Yvette, France.
Mol Microbiol. 2008 Jun;68(6):1418-27. doi: 10.1111/j.1365-2958.2008.06239.x. Epub 2008 Apr 11.
The organization of the Escherichia coli chromosome has been defined genetically as consisting of four insulated macrodomains and two less constrained regions. Here we have examined the movement of chromosomal loci by tracking fluorescent markers in time-lapse microscopy during a complete cell cycle. Analysing the positioning, the segregation pattern and the motility of markers allowed us to show that the dynamic behaviour of loci belonging to various macrodomains and less constrained regions is radically different. In macrodomains constraints on mobility are apparent whereas in non-structured regions, markers exhibited a greater motility that may explain their ability to interact with flanking macrodomains. Following replication, duplicated markers belonging to macrodomains show a colocalization step and this landmark is not apparent in non-structured regions. Chromosome segregation occurs in three steps: first, the origin-proximal half of the chromosome consisting of the Ori macrodomain and the two non-structured region segregates concomitantly in a short period of time. Second, the Right and Left macrodomains segregate progressively following the genetic map. Third, the Ter macrodomain is rapidly segregated before division, after a significant period of colocalization. Macrodomain territories defined as cellular spaces occupied by the different macrodomains can be identified.
大肠杆菌染色体的组织在遗传学上被定义为由四个绝缘的宏观结构域和两个约束较少的区域组成。在这里,我们通过在整个细胞周期的延时显微镜下跟踪荧光标记物,研究了染色体位点的移动。分析标记物的定位、分离模式和运动性,使我们能够表明,属于不同宏观结构域和约束较少区域的位点的动态行为截然不同。在宏观结构域中,对移动性的限制很明显,而在非结构化区域,标记物表现出更大的运动性,这可能解释了它们与侧翼宏观结构域相互作用的能力。复制后,属于宏观结构域的重复标记物显示出共定位步骤,而这一标志性现象在非结构化区域并不明显。染色体分离发生在三个步骤:首先,由Ori宏观结构域和两个非结构化区域组成的染色体近端一半在短时间内同时分离。其次,右和左宏观结构域按照遗传图谱逐渐分离。第三,Ter宏观结构域在经过一段显著的共定位期后,在分裂前迅速分离。可以识别被定义为不同宏观结构域占据的细胞空间的宏观结构域区域。