Regulation of DNA replication initiation by ParA is independent of parS location in Bacillus subtilis.
Regulation of DNA replication initiation by ParA is independent of parS location in Bacillus subtilis.
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DOI：
10.1099/mic.0.001259
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发表时间：
2022-10
期刊：
影响因子：
--
通讯作者：
Murray H
中科院分区：
文献类型：
--
作者：
Koh A;Strahl H;Murray H
遗传信息的复制和分离是细胞增殖所必需的。在枯草芽孢杆菌中，Par系统（ParA/Soj， ParB/Spo0J和parS）是染色体起源（oriC）区分离和DNA复制起始的适当控制所必需的。ParB结合聚集在复制原点附近的parS位点，并组装成滑动夹，与ParA相互作用，通过扩散棘轮机制驱动原点分离。作为这一动态过程的一部分，ParB刺激ParA atp酶活性，触发其从atp结合二聚体到adp结合单体的转换。除了在DNA分离中的保守作用外，ParA也是主DNA复制起始蛋白DNA的调节剂。我们假设在枯草芽孢杆菌中，Par系统靠近oriC的位置对于ParA正确调节dna是必要的。为了验证这个模型，我们构建了一系列改变了parS位点数量和位置的转基因菌株，其中许多如预期的那样干扰了染色体起源分离。与我们的假设相反，结果表明，当parS位点与oriC分离时，ParA对DNA复制起始的调控得以维持。由于单个parS位点足以适当控制ParA，因此结果与parS加载后ParB滑动夹有效调节ParA的模型一致。
Replication and segregation of the genetic information is necessary for a cell to proliferate. In Bacillus subtilis, the Par system (ParA/Soj, ParB/Spo0J and parS) is required for segregation of the chromosome origin (oriC) region and for proper control of DNA replication initiation. ParB binds parS sites clustered near the origin of replication and assembles into sliding clamps that interact with ParA to drive origin segregation through a diffusion-ratchet mechanism. As part of this dynamic process, ParB stimulates ParA ATPase activity to trigger its switch from an ATP-bound dimer to an ADP-bound monomer. In addition to its conserved role in DNA segregation, ParA is also a regulator of the master DNA replication initiation protein DnaA. We hypothesized that in B. subtilis the location of the Par system proximal to oriC would be necessary for ParA to properly regulate DnaA. To test this model, we constructed a range of genetically modified strains with altered numbers and locations of parS sites, many of which perturbed chromosome origin segregation as expected. Contrary to our hypothesis, the results show that regulation of DNA replication initiation by ParA is maintained when a parS site is separated from oriC. Because a single parS site is sufficient for proper control of ParA, the results are consistent with a model where ParA is efficiently regulated by ParB sliding clamps following loading at parS.
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