Dmowski Michal, Sitkiewicz Izabela, Ceglowski Piotr
Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawiskiego 5A, 02-106 Warsaw, Poland.
J Bacteriol. 2006 Jun;188(12):4362-72. doi: 10.1128/JB.01922-05.
High segregational stability of the streptococcal plasmid pSM19035 is achieved by the concerted action of systems involved in plasmid copy number control, multimer resolution, and postsegregational killing. In this study, we demonstrate the role of two genes, delta and omega, in plasmid stabilization by a partition mechanism. We show that these two genes can stabilize the native pSM19035 replicon as well as other theta- and sigma-type plasmids in Bacillus subtilis. In contrast to other known partition systems, in this case the two genes are transcribed separately; however, they are coregulated by the product of the parB-like gene omega. Analysis of mutants of the parA-like gene delta showed that the Walker A ATPase motif is necessary for plasmid stabilization. The ParB-like product of the omega gene binds to three regions containing repeated WATCACW heptamers, localized in the copS (regulation of plasmid copy number), delta, and omega promoter regions. We demonstrate that all three of these regions can cause partition-mediated incompatibility. Moreover, our data suggest that each of these could play the role of a centromere-like sequence. We conclude that delta and omega constitute a novel type of plasmid stabilization system.
链球菌质粒pSM19035的高分离稳定性是通过参与质粒拷贝数控制、多聚体解离和后分离杀伤的系统协同作用实现的。在本研究中,我们证明了两个基因delta和omega通过一种分配机制在质粒稳定中发挥的作用。我们表明,这两个基因可以稳定枯草芽孢杆菌中的天然pSM19035复制子以及其他θ型和σ型质粒。与其他已知的分配系统不同,在这种情况下,这两个基因是分别转录的;然而,它们由类parB基因omega的产物共同调控。对类parA基因delta突变体的分析表明,沃克A ATP酶基序对于质粒稳定是必需的。omega基因的类ParB产物与三个含有重复WATCACW七聚体的区域结合,这些区域位于copS(质粒拷贝数调控)、delta和omega启动子区域。我们证明,所有这三个区域都可导致分配介导的不相容性。此外,我们的数据表明,这些区域中的每一个都可以发挥类似着丝粒序列的作用。我们得出结论,delta和omega构成了一种新型的质粒稳定系统。