Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Ube, Japan.
PLoS One. 2011;6(6):e20063. doi: 10.1371/journal.pone.0020063. Epub 2011 Jun 10.
The molecular mechanism supporting survival at a critical high temperature (CHT) in Escherichia coli was investigated. Genome-wide screening with a single-gene knockout library provided a list of genes indispensable for growth at 47°C, called thermotolerant genes. Genes for which expression was affected by exposure to CHT were identified by DNA chip analysis. Unexpectedly, the former contents did not overlap with the latter except for dnaJ and dnaK, indicating that a specific set of non-heat shock genes is required for the organism to survive under such a severe condition. More than half of the mutants of the thermotolerant genes were found to be sensitive to H(2)O(2) at 30°C, suggesting that the mechanism of thermotolerance partially overlaps with that of oxidative stress resistance. Their encoded enzymes or proteins are related to outer membrane organization, DNA double-strand break repair, tRNA modification, protein quality control, translation control or cell division. DNA chip analyses of essential genes suggest that many of the genes encoding ribosomal proteins are down-regulated at CHT. Bioinformatics analysis and comparison with the genomic information of other microbes suggest that E. coli possesses several systems for survival at CHT. This analysis allows us to speculate that a lipopolysaccharide biosynthesis system for outer membrane organization and a sulfur-relay system for tRNA modification have been acquired by horizontal gene transfer.
我们研究了支持大肠杆菌在临界高温(CHT)下存活的分子机制。利用单基因敲除文库进行全基因组筛选,提供了一组在 47°C 下生长所必需的基因,称为耐热基因。通过 DNA 芯片分析鉴定了因暴露于 CHT 而受影响表达的基因。出乎意料的是,前者的内容除了 dnaJ 和 dnaK 之外与后者没有重叠,这表明生物体在如此恶劣的条件下存活需要一组特定的非热休克基因。耐热基因的突变体中有一半以上在 30°C 时对 H(2)O(2)敏感,这表明耐热机制部分与氧化应激抗性机制重叠。它们编码的酶或蛋白质与外膜组织、DNA 双链断裂修复、tRNA 修饰、蛋白质质量控制、翻译控制或细胞分裂有关。必需基因的 DNA 芯片分析表明,许多编码核糖体蛋白的基因在 CHT 时下调。生物信息学分析和与其他微生物的基因组信息比较表明,大肠杆菌拥有几种在 CHT 下生存的系统。这种分析使我们推测,外膜组织的脂多糖生物合成系统和 tRNA 修饰的硫中继系统是通过水平基因转移获得的。