Department of Microbiology, Jahangirnagar University, Dhaka, 1342, Bangladesh.
Micro-Nano Technology Research Center, Hosei University, Tokyo, 184-0003, Japan.
Sci China Life Sci. 2015 Sep;58(9):902-11. doi: 10.1007/s11427-015-4898-0. Epub 2015 Jul 24.
The genomic DNA of bacteria is highly compacted in a single or a few bodies known as nucleoids. Here, we have isolated Escherichia coli nucleoid by sucrose density gradient centrifugation. The sedimentation rates, structures as well as protein/ DNA composition of isolated nucleoids were then compared under various growth phases. The nucleoid structures were found to undergo changes during the cell growth; i. e., the nucleoid structure in the stationary phase was more tightly compacted than that in the exponential phase. In addition to factor for inversion stimulation (Fis), histone-like nucleoid structuring protein (H-NS), heat-unstable nucleoid protein (HU) and integration host factor (IHF) here we have identified, three new candidates of E. coli nucleoid, namely DNA-binding protein from starved cells (Dps), host factor for phage Qβ (Hfq) and suppressor of td(-) phenotype A (StpA). Our results reveal that the major components of exponential phase nucleoid are Fis, HU, H-NS, StpA and Hfq, while Dps occupies more than half of the stationary phase nucleoid. It has been known for a while that Dps is the main nucleoid-associated protein at stationary phase. From these results and the prevailing information, we propose a model for growth phase dependent changes in the structure and protein composition of nucleoid in E. coli.
细菌的基因组 DNA 高度压缩在一个或几个称为核体的结构中。在这里,我们通过蔗糖密度梯度离心法分离了大肠杆菌核体。然后比较了在不同生长阶段下分离的核体的沉降速率、结构以及蛋白质/DNA 组成。发现核体结构在细胞生长过程中发生变化;即,静止期的核体结构比指数期的更紧密。除了倒位刺激因子(Fis)、组蛋白样核体结构蛋白(H-NS)、热不稳定核体蛋白(HU)和整合宿主因子(IHF)外,我们还鉴定了三种大肠杆菌核体的新候选蛋白,即饥饿细胞中的 DNA 结合蛋白(Dps)、噬菌体 Qβ的宿主因子(Hfq)和 td(-)表型 A 的抑制因子(StpA)。我们的结果表明,指数期核体的主要成分是 Fis、HU、H-NS、StpA 和 Hfq,而 Dps 占据了静止期核体的一半以上。众所周知,Dps 是静止期核体相关蛋白的主要成分。根据这些结果和现有的信息,我们提出了一个大肠杆菌核体结构和蛋白质组成随生长阶段变化的模型。