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Genome-Wide Transcriptional Response of the Archaeon Thermococcus gammatolerans to Cadmium

机译:古细菌嗜热球菌对镉的全基因组转录反应。

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摘要

Thermococcus gammatolerans, the most radioresistant archaeon known to date, is an anaerobic and hyperthermophilic sulfur-reducing organism living in deep-sea hydrothermal vents. Knowledge of mechanisms underlying archaeal metal tolerance in such metal-rich ecosystem is still poorly documented. We showed that T. gammatolerans exhibits high resistance to cadmium (Cd), cobalt (Co) and zinc (Zn), a weaker tolerance to nickel (Ni), copper (Cu) and arsenate (AsO4) and that cells exposed to 1 mM Cd exhibit a cellular Cd concentration of 67 µM. A time-dependent transcriptomic analysis using microarrays was performed at a non-toxic (100 µM) and a toxic (1 mM) Cd dose. The reliability of microarray data was strengthened by real time RT-PCR validations. Altogether, 114 Cd responsive genes were revealed and a substantial subset of genes is related to metal homeostasis, drug detoxification, re-oxidization of cofactors and ATP production. This first genome-wide expression profiling study of archaeal cells challenged with Cd showed that T. gammatolerans withstands induced stress through pathways observed in both prokaryotes and eukaryotes but also through new and original strategies. T. gammatolerans cells challenged with 1 mM Cd basically promote: 1) the induction of several transporter/permease encoding genes, probably to detoxify the cell; 2) the upregulation of Fe transporters encoding genes to likely compensate Cd damages in iron-containing proteins; 3) the induction of membrane-bound hydrogenase (Mbh) and membrane-bound hydrogenlyase (Mhy2) subunits encoding genes involved in recycling reduced cofactors and/or in proton translocation for energy production. By contrast to other organisms, redox homeostasis genes appear constitutively expressed and only a few genes encoding DNA repair proteins are regulated. We compared the expression of 27 Cd responsive genes in other stress conditions (Zn, Ni, heat shock, γ-rays), and showed that the Cd transcriptional pattern is comparable to other metal stress transcriptional responses (Cd, Zn, Ni) but not to a general stress response.
机译:高温球菌是迄今为止已知的最耐辐射的古细菌,是生活在深海热液喷口中的一种厌氧和超嗜热的硫还原生物。在这种富含金属的生态系统中,对古细菌金属耐受性的潜在机制的了解仍然很少。我们表明,T。gammatolerans表现出对镉(Cd),钴(Co)和锌(Zn)的高抗性,对镍(Ni),铜(Cu)和砷酸(AsO4)的较弱耐受性,并且细胞暴露于1 mM Cd的细胞Cd浓度为67 µM。使用微阵列进行了随时间变化的转录组分析,以无毒(100 µM)和有毒(1 mM)镉剂量进行。实时RT-PCR验证增强了微阵列数据的可靠性。总共揭示了114个Cd响应基因,其中大部分基因与金属稳态,药物解毒,辅因子的再氧化和ATP产生有关。这项首次对Cd攻击的古细菌细胞进行全基因组表达分析的研究表明,γ耐受的T. glertolerans可以通过原核生物和真核生物中观察到的途径耐受诱导的应激,而且还可以通过新的和原始的策略进行耐受。用1 mM Cd攻击的T. gammatolerans细胞基本上可以促进:1)诱导几种转运蛋白/通透酶编码基因,可能使细胞解毒; 2)铁转运蛋白编码基因的上调可能补偿含铁蛋白质中的镉损伤; 3)诱导与膜结合的加氢酶(Mbh)和膜结合的加氢酶(Mhy2)亚基的编码,这些基因参与回收减少的辅因子和/或质子转运以产生能量。与其他生物相反,氧化还原稳态基因似乎是组成型表达的,只有少数编码DNA修复蛋白的基因受到调控。我们比较了27种Cd响应基因在其他胁迫条件下(锌,镍,热激,γ射线)的表达,结果表明Cd转录模式与其他金属胁迫转录响应(Cd,Zn,Ni)相当,但不对一般的压力反应。

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