Department of Life Science and Medical Bioscience, Waseda University, Tokyo, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan.
Microbiologyopen. 2017 Oct;6(5). doi: 10.1002/mbo3.501. Epub 2017 Jul 3.
Microbes are known to withstand environmental stresses by using chromosomal toxin-antitoxin systems. MazEF is one of the most extensively studied toxin-antitoxin systems. In stressful environments, MazF toxins modulate translation by cleaving single-stranded RNAs in a sequence-specific fashion. Previously, a chromosomal gene located at DR0417 in Deinococcus radiodurans was predicted to code for a MazF endoribonuclease (MazF ); however, its function remains unclear. In the present study, we characterized the molecular function of MazF . Analysis of MazF -cleaved RNA sites using modified massively parallel sequencing revealed a unique 4-nt motif, UACA, as a potential cleavage pattern. The activity of MazF was also assessed in a real-time fluorometric assay, which revealed that MazF strictly recognizes the unique tetrad UACA. This sequence specificity may allow D. radiodurans to alter its translation profile and survive under stressful conditions.
微生物被认为可以通过使用染色体毒素-抗毒素系统来耐受环境压力。 MazEF 是研究最广泛的毒素-抗毒素系统之一。在应激环境中,MazF 毒素通过序列特异性方式切割单链 RNA 来调节翻译。先前,在 Deinococcus radiodurans 中位于 DR0417 的染色体基因被预测编码为 MazF 内切核糖核酸酶(MazF);然而,其功能仍不清楚。在本研究中,我们表征了 MazF 的分子功能。使用改良的大规模平行测序分析 MazF 切割的 RNA 位点揭示了一个独特的 4 个核苷酸基序 UACA,作为潜在的切割模式。MazF 的活性也在实时荧光测定中进行了评估,结果表明 MazF 严格识别独特的四联体 UACA。这种序列特异性可能使 Deinococcus radiodurans 能够改变其翻译谱并在应激条件下存活。