Choi Wonho, Yamaguchi Yoshihiro, Lee Jae-Woo, Jang Kyung-Min, Inouye Masayori, Kim Sung-Gun, Yoon Min-Ho, Park Jung-Ho
Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, South Korea.
Department of Bio-Environmental Chemistry, College of Agriculture and Life Sciences, Chungnam National University, Yuseong-gu, South Korea.
FEBS Lett. 2017 Jul;591(13):1853-1861. doi: 10.1002/1873-3468.12705. Epub 2017 Jun 26.
Many bacteria have toxin-antitoxin (TA) systems, where toxin gene expression inhibits their own cell growth. mRNA is one of the well-known targets of the toxins in the type II toxin-antitoxin systems. Here, we examined the ribosome dependency of the endoribonuclease activity of YhaV, one of the toxins in type II TA systems, on mRNA in vitro and in vivo. A polysome profiling assay revealed that YhaV is bound to the 70S ribosomes and 50S ribosomal subunits. Moreover, we found that while YhaV cleaves ompF and lpp mRNAs in a translation-dependent manner, they did not cleave the 5' untranslated region in primer extension experiments. From these results, we conclude that YhaV is a ribosome-dependent toxin that cleaves mRNA in a translation-dependent manner.
许多细菌拥有毒素-抗毒素(TA)系统,其中毒素基因的表达会抑制其自身细胞生长。mRNA是II型毒素-抗毒素系统中毒素的已知靶标之一。在此,我们在体外和体内研究了II型TA系统中的毒素之一YhaV的核糖核酸内切酶活性对mRNA的核糖体依赖性。多核糖体谱分析显示YhaV与70S核糖体和50S核糖体亚基结合。此外,我们发现虽然YhaV以翻译依赖性方式切割ompF和lpp mRNA,但在引物延伸实验中它们并未切割5'非翻译区。从这些结果中,我们得出结论,YhaV是一种核糖体依赖性毒素,以翻译依赖性方式切割mRNA。