Laboratoire de Biochimie et de Biologie Moléculaire, Faculté des sciences de Bizerte, 7021 Zarzouna, Tunisia.
Plasmid. 2011 Jan;65(1):15-9. doi: 10.1016/j.plasmid.2010.09.003. Epub 2010 Sep 25.
The pBR322 plasmid origin replication and oriC show similar responses to adenine methylation. Both are subject to sequestration by membrane fractions. In fact, like the host origin oriC, the RNA II promoter region of pBR322 is regulated by methylation of three GATC adenine methylation sites. The SeqA gene product acts in the negative control of oriC by sequestration. We suggest that the role of SeqA protein in sequestration is similar to oriC region DNA. Hence, SeqA recognize the methylation state of the pBR322RNA II promoter region by direct DNA binding in vitro. Using the pOC42 plasmid, we show that SeqA binds exclusively to the hemimethylated form of the replication origin of the pBR322 plasmid. In addition, we suggested that the SeqA protein could modulate periodically the initiation of replication of the pBR322 plasmid. The later could be fixed by its origin sequence, on a hemimethylated state, during the initiation of the replication.
pBR322 质粒复制原点和 oriC 对腺嘌呤甲基化有相似的反应。两者都受到膜部分的隔离。事实上,与宿主起源 oriC 一样,pBR322 的 RNA II 启动子区域受到三个 GATC 腺嘌呤甲基化位点的甲基化调控。SeqA 基因产物通过隔离在负调控 oriC。我们认为 SeqA 蛋白在隔离中的作用与 oriC 区域 DNA 相似。因此,SeqA 可以通过体外直接 DNA 结合来识别 pBR322RNA II 启动子区域的甲基化状态。使用 pOC42 质粒,我们表明 SeqA 仅结合于 pBR322 质粒复制原点的半甲基化形式。此外,我们认为 SeqA 蛋白可以周期性地调节 pBR322 质粒复制的起始。后者可以通过其起源序列固定在半甲基化状态,在复制起始时。