Lab. of Biochemistry and Molecular Biology, Center for Cancer Research, NCI, 37 Convent Drive, Rm. 6044, NIH, Bethesda, MD 20892-4260, USA.
J Bacteriol. 2011 Apr;193(7):1504-14. doi: 10.1128/JB.01067-10. Epub 2011 Jan 21.
Vibrio cholerae carries homologs of plasmid-borne parA and parB genes on both of its chromosomes. The par genes help to segregate many plasmids and chromosomes. Here we have studied the par genes of V. cholerae chromosome I. Earlier studies suggested that ParBI binds to the centromeric site parSI near the origin of replication (oriI), and parSI-ParBI complexes are placed at the cell poles by ParAI. Deletion of parAI and parSI caused the origin-proximal DNA to be less polar. Here we found that deletion of parBI also resulted in a less polar localization of oriI. However, unlike the deletion of parAI, the deletion of parBI increased the oriI number. Replication was normal when both parAI and parBI were deleted, suggesting that ParBI mediates its action through ParAI. Overexpression of ParAI in a parABI-deleted strain also increased the DNA content. The results are similar to those found for Bacillus subtilis, where ParA (Soj) stimulates replication and this activity is repressed by ParB (SpoOJ). As in B. subtilis, the stimulation of replication most likely involves the replication initiator DnaA. Our results indicate that control of chromosomal DNA replication is an additional function of chromosomal par genes conserved across the Gram-positive/Gram-negative divide.
霍乱弧菌的两条染色体上都携带质粒携带的 parA 和 parB 基因的同源物。这些 par 基因有助于分离许多质粒和染色体。在这里,我们研究了霍乱弧菌染色体 I 的 par 基因。早期的研究表明,ParBI 结合到复制起点(oriI)附近的着丝粒位点 parSI,并且 parSI-ParBI 复合物通过 ParAI 被放置在细胞两极。parAI 和 parSI 的缺失导致起始近端 DNA 的极性降低。在这里,我们发现 parBI 的缺失也导致 oriI 的定位变得不那么极性。然而,与缺失 parAI 不同的是,parBI 的缺失增加了 oriI 的数量。当同时缺失 parAI 和 parBI 时,复制是正常的,这表明 ParBI 通过 ParAI 发挥其作用。在 parABI 缺失的菌株中过表达 ParAI 也增加了 DNA 含量。结果与枯草芽孢杆菌相似,其中 ParA(Soj)刺激复制,而这种活性被 ParB(SpoOJ)抑制。与枯草芽孢杆菌一样,复制的刺激很可能涉及复制起始因子 DnaA。我们的结果表明,染色体 DNA 复制的控制是革兰氏阳性/革兰氏阴性分裂中保守的染色体 par 基因的另一个功能。