Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Medical School, Newcastle University, Newcastle, UK.
Mol Microbiol. 2010 Jan;75(2):333-48. doi: 10.1111/j.1365-2958.2009.06969.x. Epub 2009 Nov 25.
Prokaryotic toxin - antitoxin (TA) loci encode mRNA interferases that inhibit translation, either by cleaving mRNA codons at the ribosomal A site or by cleaving any RNA site-specifically. So far, seven mRNA interferases of Escherichia coli have been identified, four of which cleave mRNA by a translation-dependent mechanism. Here, we experimentally confirmed the presence of three novel TA loci in E. coli. We found that the yafNO, higBA (ygjNM) and ygiUT loci encode mRNA interferases related to RelE. YafO and HigB cleaved translated mRNA only, while YgiU cleaved RNA site-specifically at GC[A/U], independently of translation. Thus, YgiU is the first RelE-related mRNA interferase that cleaves mRNA independently of translation, in vivo. All three loci were induced by amino acid starvation, and inhibition of translation although to different degrees. Carbon starvation induced only two of the loci. The yafNO locus was induced by DNA damage, but the transcription originated from the dinB promoter. Thus, our results showed that the different TA loci responded differentially to environmental stresses. Induction of the three loci depended on Lon protease that may sense the environmental stresses and activate TA loci by cleavage of the antitoxins. Transcription of the three TA operons was autoregulated by the antitoxins.
原核生物毒素-抗毒素(TA)基因座编码的 mRNA 干扰物通过核糖体 A 位切割 mRNA 密码子或通过特异性切割任何 RNA 位点来抑制翻译。迄今为止,已在大肠杆菌中鉴定出七种 mRNA 干扰物,其中四种通过依赖翻译的机制切割 mRNA。在这里,我们通过实验证实了大肠杆菌中存在三个新的 TA 基因座。我们发现 yafNO、higBA(ygjNM)和 ygiUT 基因座编码与 RelE 相关的 mRNA 干扰物。YafO 和 HigB 仅切割翻译的 mRNA,而 YgiU 则独立于翻译在 GC[A/U]处特异性切割 RNA。因此,YgiU 是第一个在体内独立于翻译切割 mRNA 的 RelE 相关 mRNA 干扰酶。所有三个基因座都被氨基酸饥饿诱导,尽管翻译受到不同程度的抑制。碳饥饿仅诱导其中两个基因座。yafNO 基因座由 DNA 损伤诱导,但转录起始于 dinB 启动子。因此,我们的结果表明,不同的 TA 基因座对环境应激有不同的反应。三个基因座的诱导依赖于 Lon 蛋白酶,Lon 蛋白酶可能感知环境应激并通过切割抗毒素来激活 TA 基因座。三个 TA 操纵子的转录受抗毒素的自身调控。