Department of Frontier Bioscience, Hosei University, Koganei, Tokyo 184-8584, Japan.
Nucleic Acids Res. 2010 Jun;38(11):3605-18. doi: 10.1093/nar/gkq077. Epub 2010 Feb 15.
A systematic search was performed for DNA-binding sequences of YgiP, an uncharacterized transcription factor of Escherichia coli, by using the Genomic SELEX. A total of 688 YgiP-binding loci were identified after genome-wide profiling of SELEX fragments with a high-density microarray (SELEX-chip). Gel shift and DNase-I footprinting assays indicated that YgiP binds to multiple sites along DNA probes with a consensus GTTNATT sequence. Atomic force microscope observation indicated that at low concentrations, YgiP associates at various sites on DNA probes, but at high concentrations, YgiP covers the entire DNA surface supposedly through protein-protein contact. The intracellular concentration of YgiP is very low in growing E. coli cells under aerobic conditions, but increases more than 100-fold to the level as high as the major nucleoid proteins under anaerobic conditions. An E. coli mutant lacking ygiP showed retarded growth under anaerobic conditions. High abundance and large number of binding sites together indicate that YgiP is a nucleoid-associated protein with both architectural and regulatory roles as the nucleoid proteins Fis and IHF. We then propose that YgiP is a novel nucleoid protein of E. coli under anaerobiosis and propose to rename it Dan (DNA-binding protein under anaerobic conditions).
通过基因组 SELEX 对大肠杆菌中未鉴定的转录因子 YgiP 的 DNA 结合序列进行了系统搜索。在用高密度微阵列(SELEX-chip)对 SELEX 片段进行全基因组分析后,共鉴定出 688 个 YgiP 结合位点。凝胶迁移和 DNase-I 足迹分析表明,YgiP 结合到 DNA 探针上的多个位点,其共有序列为 GTTNATT。原子力显微镜观察表明,在低浓度下,YgiP 与 DNA 探针上的各种位点结合,但在高浓度下,YgiP 通过蛋白-蛋白接触覆盖整个 DNA 表面。在有氧条件下生长的大肠杆菌细胞中,YgiP 的细胞内浓度非常低,但在无氧条件下,其浓度增加 100 多倍,达到与主要核区蛋白一样高的水平。缺乏 ygiP 的大肠杆菌突变体在无氧条件下生长缓慢。高丰度和大量的结合位点共同表明,YgiP 是一种核区相关蛋白,具有作为核区蛋白 Fis 和 IHF 的结构和调节作用。因此,我们提出 YgiP 是大肠杆菌在无氧条件下的一种新的核区蛋白,并提议将其重新命名为 Dan(缺氧条件下的 DNA 结合蛋白)。