Ohniwa Ryosuke L, Morikawa Kazuya, Kim Joongbaek, Ohta Toshiko, Ishihama Akira, Wada Chieko, Takeyasu Kunio
Laboratory of Plasma Membrane and Nuclear Signaling, Kyoto University Graduate School of Biostudies, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, Japan.
EMBO J. 2006 Nov 29;25(23):5591-602. doi: 10.1038/sj.emboj.7601414. Epub 2006 Nov 9.
In bacteria, Dps is one of the critical proteins to build up a condensed nucleoid in response to the environmental stresses. In this study, we found that the expression of Dps and the nucleoid condensation was not simply correlated in Escherichia coli, and that Fis, which is an E. coli (gamma-Proteobacteria)-specific nucleoid protein, interfered with the Dps-dependent nucleoid condensation. Atomic force microscopy and Northern blot analyses indicated that the inhibitory effect of Fis was due to the repression of the expression of Topoismerase I (Topo I) and DNA gyrase. In the Deltafis strain, both topA and gyrA/B genes were found to be upregulated. Overexpression of Topo I and DNA gyrase enhanced the nucleoid condensation in the presence of Dps. DNA-topology assays using the cell extract showed that the extracts from the Deltafis and Topo I-/DNA gyrase-overexpressing strains, but not the wild-type extract, shifted the population toward relaxed forms. These results indicate that the topology of DNA is dynamically transmutable and that the topology control is important for Dps-induced nucleoid condensation.
在细菌中,Dps是一种在应对环境压力时构建浓缩类核的关键蛋白质。在本研究中,我们发现,在大肠杆菌中,Dps的表达与类核浓缩并非简单相关,并且Fis(一种大肠杆菌(γ-变形菌纲)特异性类核蛋白)会干扰Dps依赖的类核浓缩。原子力显微镜和Northern印迹分析表明,Fis的抑制作用是由于拓扑异构酶I(Topo I)和DNA促旋酶表达受抑制所致。在Δfis菌株中,发现topA和gyrA/B基因均上调。Topo I和DNA促旋酶的过表达在有Dps存在时增强了类核浓缩。使用细胞提取物进行的DNA拓扑分析表明,来自Δfis菌株和Topo I-/DNA促旋酶过表达菌株的提取物,而非野生型提取物,使群体向松弛形式转变。这些结果表明,DNA的拓扑结构是动态可变的,并且拓扑控制对Dps诱导的类核浓缩很重要。