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首页> 外文期刊>Journal of bacteriology >Multicopy Plasmids Affect Replisome Positioning in Bacillus subtilis
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Multicopy Plasmids Affect Replisome Positioning in Bacillus subtilis

机译：多拷贝质粒影响枯草芽孢杆菌中的复制体定位。

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The DNA replication machinery, various regions of the chromosome, and some plasmids occupy characteristic subcellular positions in bacterial cells. We visualized the location of a multicopy plasmid, pHP13, in living cells of Bacillus subtilis using an array of lac operators and LacI-green fluorescent protein (GFP). In the majority of cells, plasmids appeared to be highly mobile and randomly distributed. In a small fraction of cells, there appeared to be clusters of plasmids located predominantly at or near a cell pole. We also monitored the effects of the presence of multicopy plasmids on the position of DNA polymerase using a fusion of a subunit of DNA polymerase to GFP. Many of the plasmid-containing cells had extra foci of the replisome, and these were often found at uncharacteristic locations in the cell. Some of the replisome foci were dynamic and highly mobile, similar to what was observed for the plasmid. In contrast, replisome foci in plasmid-free cells were relatively stationary. Our results indicate that in B. subtilis, plasmid-associated replisomes are recruited to the subcellular position of the plasmid. Extending this notion to the chromosome, we postulated that the subcellular position of the chromosomally associated replisome is established by the subcellular location of oriC at the time of initiation of replication.
机译：DNA复制机制，染色体的各个区域和某些质粒在细菌细胞中占据特征性的亚细胞位置。我们使用 lac 操纵子和LacI-绿色荧光蛋白（GFP）阵列，在枯草芽孢杆菌的活细胞中可视化了多拷贝质粒pHP13的位置。在大多数细胞中，质粒似乎具有高度的移动性并随机分布。在一小部分细胞中，似乎有大量质粒簇位于细胞极或细胞极附近。我们还使用DNA聚合酶亚基与GFP的融合，监测了多拷贝质粒的存在对DNA聚合酶位置的影响。许多含有质粒的细胞具有额外的复制体灶，并且通常在细胞中不常见的位置发现它们。一些复制体焦点是动态的并且高度可移动的，类似于在质粒中观察到的。相反，无质粒细胞中的复制子灶相对固定。我们的结果表明在 B中。枯草杆菌，与质粒相关的复制体被募集到质粒的亚细胞位置。将此概念扩展到染色体上，我们推测染色体相关复制体的亚细胞位置是由复制开始时 oriC 的亚细胞位置确定的。

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