tmRNA regulates synthesis of the ArfA ribosome rescue factor.

tmRNA regulates synthesis of the ArfA ribosome rescue factor.
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DOI:
10.1111/j.1365-2958.2011.07638.x
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发表时间:
2011-06
影响因子:
3.6
通讯作者:
Hayes CS
Hayes CS
中科院分区:
生物学2区
文献类型:
--
作者:
Garza-Sánchez F;Schaub RE;Janssen BD;Hayes CS

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缺少框内终止密码子的mRNA的翻译会导致转录本3‘端的核糖体停滞。在细菌中,tmRNA质量控制系统回收这些停滞不前的核糖体,并标记不完整的新生链供降解。虽然在真细菌中普遍存在,但编码tmRNA的ssrA基因对大肠杆菌和其他模式细菌物种的生存并不是必不可少的。ARFA(YhdL)是一种tmRNA非依赖性核糖体拯救的介体,对大肠杆菌Δ单链RNA突变体的生存是必不可少的。在这里,我们证明了ARFA是由截短的mRNA合成的,因此受到tmRNA标签活性的调节。RNaseIII切割ARFA编码序列中的发夹结构,产生缺乏终止密码子的转录本。在没有tmRNA标签的情况下,截短的Arfa链从核糖体中释放出来。截短的ArfAΔ18蛋白(缺少18个C末端残基)在核糖体拯救中具有功能,当从ArfA基因座表达时,它支持ΔSSrA细胞的生存。其他蛋白质细菌的arfa基因也编码发夹,来自迪克雅氏杆菌和鼠伤寒沙门氏菌的转录本在大肠杆菌中表达时被RNaseIII切割。因此,从截短的mRNA合成ARFA似乎是调节替代核糖体拯救活性的一般机制。
Translation of mRNA lacking an in-frame stop codon leads to ribosome arrest at the 3´-end of the transcript. In bacteria, the tmRNA quality control system recycles these stalled ribosomes and tags the incomplete nascent chains for degradation. Although ubiquitous in eubacteria, the ssrA gene encoding tmRNA is not essential for the viability of Escherichia coli and other model bacterial species. ArfA (YhdL) is a mediator of tmRNA-independent ribosome rescue that is essential for the viability of E. coli ΔssrA mutants. Here, we demonstrate that ArfA is synthesized from truncated mRNA and therefore regulated by tmRNA tagging activity. RNase III cleaves a hairpin structure within the arfA coding sequence to produce transcripts that lack stop codons. In the absence of tmRNA tagging, truncated ArfA chains are released from the ribosome. The truncated ArfAΔ18 protein (which lacks 18 C-terminal residues) is functional in ribosome rescue and supports ΔssrA cell viability when expressed from the arfA locus. Other proteobacterial arfA genes also encode hairpins, and transcripts from Dickeya dadantii and Salmonella typhimurium are cleaved by RNase III when expressed in E. coli. Thus, synthesis of ArfA from truncated mRNA appears to be a general mechanism to regulate alternative ribosome rescue activity.
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