Gras Konrad, Fange David, Elf Johan
Dept. of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
Nat Commun. 2024 Jul 17;15(1):6018. doi: 10.1038/s41467-024-50047-z.
In Escherichia coli, it is debated whether the two replisomes move independently along the two chromosome arms during replication or if they remain spatially confined. Here, we use high-throughput fluorescence microscopy to simultaneously determine the location and short-time-scale (1 s) movement of the replisome and a chromosomal locus throughout the cell cycle. The assay is performed for several loci. We find that (i) the two replisomes are confined to a region of ~250 nm and ~120 nm along the cell's long and short axis, respectively, (ii) the chromosomal loci move to and through this region sequentially based on their distance from the origin of replication, and (iii) when a locus is being replicated, its short time-scale movement slows down. This behavior is the same at different growth rates. In conclusion, our data supports a model with DNA moving towards spatially confined replisomes at replication.
在大肠杆菌中,关于两个复制体在复制过程中是沿着两条染色体臂独立移动还是在空间上受到限制存在争议。在这里,我们使用高通量荧光显微镜在整个细胞周期中同时确定复制体和一个染色体位点的位置以及短时间尺度(1秒)的移动。对几个位点进行了该测定。我们发现:(i)两个复制体分别沿着细胞的长轴和短轴被限制在约250纳米和约120纳米的区域内;(ii)染色体位点根据它们与复制起点的距离依次移动到该区域并穿过该区域;(iii)当一个位点正在被复制时,其短时间尺度的移动会减慢。这种行为在不同的生长速率下是相同的。总之,我们的数据支持一种模型,即在复制时DNA朝着空间受限的复制体移动。