Importance of diphthamide modified EF2 for translational accuracy and competitive cell growth in yeast.
Importance of diphthamide modified EF2 for translational accuracy and competitive cell growth in yeast.
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DOI：
10.1371/journal.pone.0205870
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Schaffrath R
中科院分区：
文献类型：
--
作者：
Hawer H;Ütkür K;Arend M;Mayer K;Adrian L;Brinkmann U;Schaffrath R
在真核生物中，翻译延伸因子2(EF2)中的不变组氨酸(酵母中为His-699)残基用敌草胺修饰涉及一条由DPH1-DPH7基因网络编码的保守途径。白喉毒素和相关的致死性ADP核糖基酶是白喉毒素的靶标，它们通过在mRNA翻译和蛋白质生物合成过程中灭活EF2的基本转位酶功能来集体杀死细胞。尽管这一概念强调了敌百虫胺的病理重要性，但包括我们自己的细胞在内的细胞为什么需要EF2携带它的确切原因尚不清楚。从发芽酵母中挖掘合成遗传阵列(SGA)图谱，揭示了EF2(EFT1-EFT2)和敌草胺(DPH1-DPH7)基因缺失之间的负相互作用。根据这些相关性，我们在这里证实了EF2上失去双苯二甲胺修饰(DphΔ)和EF2供应不足(Ef2Δ)导致了复合突变体(DphΔEf2Δ)中的合成生长表型。它们反映了在标准和热和/或化学应激条件下对细胞性能的负面干扰、细胞生长速度和倍增时间、竞争适应性、在TOR抑制剂(雷帕霉素、咖啡因)和翻译指示药物(潮霉素、山奈霉素)存在下的细胞存活率。在缺乏EF2可修饰池的突变体(DphΔ，DphEF2mRNA)中，细胞毒性Dph5过表达显著抑制了对EF2抑制的耐受性，增加了核糖体-1帧移错误，我们的数据表明，敌百胺在Δ翻译过程中对EF2转位功能的保真度是重要的。
In eukaryotes, the modification of an invariant histidine (His-699 in yeast) residue in translation elongation factor 2 (EF2) with diphthamide involves a conserved pathway encoded by the DPH1-DPH7 gene network. Diphthamide is the target for diphtheria toxin and related lethal ADP ribosylases, which collectively kill cells by inactivating the essential translocase function of EF2 during mRNA translation and protein biosynthesis. Although this notion emphasizes the pathological importance of diphthamide, precisely why cells including our own require EF2 to carry it, is unclear. Mining the synthetic genetic array (SGA) landscape from the budding yeast Saccharomyces cerevisiae has revealed negative interactions between EF2 (EFT1-EFT2) and diphthamide (DPH1-DPH7) gene deletions. In line with these correlations, we confirm in here that loss of diphthamide modification (dphΔ) on EF2 combined with EF2 undersupply (eft2Δ) causes synthetic growth phenotypes in the composite mutant (dphΔ eft2Δ). These reflect negative interference with cell performance under standard as well as thermal and/or chemical stress conditions, cell growth rates and doubling times, competitive fitness, cell viability in the presence of TOR inhibitors (rapamycin, caffeine) and translation indicator drugs (hygromycin, anisomycin). Together with significantly suppressed tolerance towards EF2 inhibition by cytotoxic DPH5 overexpression and increased ribosomal -1 frame-shift errors in mutants lacking modifiable pools of EF2 (dphΔ, dphΔ eft2Δ), our data indicate that diphthamide is important for the fidelity of the EF2 translocation function during mRNA translation.
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