Cass Julie A, Kuwada Nathan J, Traxler Beth, Wiggins Paul A
Departments of Physics, Bioengineering, and Microbiology, University of Washington, Seattle, Washington.
Departments of Physics, Bioengineering, and Microbiology, University of Washington, Seattle, Washington.
Biophys J. 2016 Jun 21;110(12):2597-2609. doi: 10.1016/j.bpj.2016.04.046.
The structure of the Escherichia coli chromosome is inherently dynamic over the duration of the cell cycle. Genetic loci undergo both stochastic motion around their initial positions and directed motion to opposite poles of the rod-shaped cell during segregation. We developed a quantitative method to characterize cell-cycle dynamics of the E. coli chromosome to probe the chromosomal steady-state mobility and segregation process. By tracking fluorescently labeled chromosomal loci in thousands of cells throughout the entire cell cycle, our method allows for the statistical analysis of locus position and motion, the step-size distribution for movement during segregation, and the locus drift velocity. The robust statistics of our detailed analysis of the wild-type E. coli nucleoid allow us to observe loci moving toward midcell before segregation occurs, consistent with a replication factory model. Then, as segregation initiates, we perform a detailed characterization of the average segregation velocity of loci. Contrary to origin-centric models of segregation, which predict distinct dynamics for oriC-proximal versus oriC-distal loci, we find that the dynamics of loci were universal and independent of genetic position.
在细胞周期的持续时间内,大肠杆菌染色体的结构本质上是动态的。遗传位点在其初始位置周围进行随机运动,并在分离过程中向杆状细胞的两极进行定向运动。我们开发了一种定量方法来表征大肠杆菌染色体的细胞周期动态,以探究染色体的稳态迁移率和分离过程。通过在整个细胞周期中跟踪数千个细胞中荧光标记的染色体位点,我们的方法允许对位点位置和运动、分离过程中运动的步长分布以及位点漂移速度进行统计分析。我们对野生型大肠杆菌类核的详细分析的稳健统计数据使我们能够观察到在分离发生之前位点向细胞中部移动,这与复制工厂模型一致。然后,随着分离开始,我们对位点的平均分离速度进行了详细表征。与以原点为中心的分离模型相反,该模型预测oriC近端与oriC远端位点有不同的动态,我们发现位点的动态是普遍的,且与基因位置无关。