Kim Joongbaek, Yoshimura Shige H, Hizume Kohji, Ohniwa Ryosuke L, Ishihama Akira, Takeyasu Kunio
Laboratory of Plasma Membrane and Nuclear Signaling, Graduate School of Biostudies, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
Nucleic Acids Res. 2004 Apr 1;32(6):1982-92. doi: 10.1093/nar/gkh512. Print 2004.
A small container of several to a few hundred microm3 (i.e. bacterial cells and eukaryotic nuclei) contains extremely long genomic DNA (i.e. mm and m long, respectively) in a highly organized fashion. To understand how such genomic architecture could be achieved, Escherichia coli nucleoids were subjected to structural analyses under atomic force microscopy, and found to change their structure dynamically during cell growth, i.e. the nucleoid structure in the stationary phase was more tightly compacted than in the log phase. However, in both log and stationary phases, a fundamental fibrous structure with a diameter of approximately 80 nm was found. In addition to this '80 nm fiber', a thinner '40 nm fiber' and a higher order 'loop' structure were identified in the log phase nucleoid. In the later growth phases, the nucleoid turned into a 'coral reef structure' that also possessed the 80 nm fiber units, and, finally, into a 'tightly compacted nucleoid' that was stable in a mild lysis buffer. Mutant analysis demonstrated that these tight compactions of the nucleoid required a protein, Dps. From these results and previously available information, we propose a structural model of the E.coli nucleoid.
一个体积为几到几百立方微米的小容器(即细菌细胞和真核细胞核)以高度有序的方式容纳着极其长的基因组DNA(分别为毫米和米级长度)。为了理解这种基因组结构是如何形成的,对大肠杆菌类核进行了原子力显微镜下的结构分析,发现其在细胞生长过程中结构会动态变化,即稳定期的类核结构比对数期更紧密。然而,在对数期和稳定期,都发现了一种直径约为80纳米的基本纤维结构。除了这种“80纳米纤维”外,在对数期类核中还鉴定出了更细的“40纳米纤维”和更高层次的“环状”结构。在生长后期,类核变成了同样具有80纳米纤维单元的“珊瑚礁结构”,最终变成了在温和裂解缓冲液中稳定的“紧密压缩类核”。突变分析表明,类核的这些紧密压缩需要一种蛋白质,即Dps。根据这些结果和先前已有的信息,我们提出了大肠杆菌类核的结构模型。