Chiu Chung-Min, Manzoor Susan E, Batt Sarah M, Muntaha Sidra tul, Bingle Lewis E H, Thomas Christopher M
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Plasmid. 2008 May;59(3):163-75. doi: 10.1016/j.plasmid.2008.02.001.
The ParB family partitioning protein, KorB, of plasmid RK2 is central to a regulatory network coordinating replication, maintenance and transfer genes. Previous immunofluorescence microscopy indicated that the majority of KorB is localized in plasmid foci. The 12 identified KorB binding sites on RK2 are differentiated by: position relative to promoters; binding strength; and cooperativity with other repressors and so the distribution of KorB may be sequestered around a sub-set of sites. However, chromatin immunoprecipitation analysis showed that while RK2 DNA molecules appear to sequester KorB to create a higher local concentration, cooperativity between DNA binding proteins does not result in major differences in binding site occupancy. Thus under steady state conditions all operators are close to fully occupied and this correlates with gene expression on the plasmid being highly repressed.
质粒RK2的ParB家族分区蛋白KorB是协调复制、维持和转移基因的调控网络的核心。先前的免疫荧光显微镜检查表明,大多数KorB定位于质粒焦点。在RK2上已鉴定出的12个KorB结合位点的区别在于:相对于启动子的位置;结合强度;以及与其他阻遏物的协同作用,因此KorB的分布可能被隔离在一部分位点周围。然而,染色质免疫沉淀分析表明,虽然RK2 DNA分子似乎隔离KorB以产生更高的局部浓度,但DNA结合蛋白之间的协同作用不会导致结合位点占有率的重大差异。因此,在稳态条件下,所有操纵基因几乎都被完全占据,这与质粒上的基因表达受到高度抑制相关。