Department of Biology and Geosciences, Graduate School of Science, Osaka City University, Osaka, Japan.
Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Republic of Korea.
J Mol Microbiol Biotechnol. 2018;28(6):269-280. doi: 10.1159/000497146. Epub 2019 Mar 20.
MazF is a sequence-specific endoribonuclease that is widely conserved in bacteria and archaea. Here, we found an MazF homologue (MazF-lp; LPO-p0114) in Legionella pneumophila. The mazF-lp gene overlaps 14 base pairs with the upstream gene mazE-lp (MazE-lp; LPO-p0115). The induction of mazF-lp caused cell growth arrest, while mazE-lp co-induction recovered cell growth in Escherichia coli. In vivo and in vitro primer extension experiments showed that MazF-lp is a sequence-specific endoribonuclease cleaving RNA at AACU. The endoribonuclease activity of purified MazF-lp was inhibited by purified MazE-lp. We found that MazE-lp and the MazEF-lp complex specifically bind to the palindromic sequence present in the 5'-untranslated region of the mazEF-lp operon. MazE-lp and MazEF-lp both likely function as a repressor for the mazEF-lp operon and for other genes, including icmR, whose gene product functions as a secretion chaperone for the IcmQ pore-forming protein, by specifically binding to the palindromic sequence in 5'-UTR of these genes.
MazF 是一种在细菌和古菌中广泛保守的序列特异性内切核糖核酸酶。在这里,我们在嗜肺军团菌中发现了一种 MazF 同源物(MazF-lp;LPO-p0114)。mazF-lp 基因与上游基因 mazE-lp(MazE-lp;LPO-p0115)重叠 14 个碱基对。mazF-lp 的诱导导致细胞生长停滞,而 mazE-lp 的共诱导恢复了大肠杆菌中的细胞生长。体内和体外引物延伸实验表明,MazF-lp 是一种序列特异性内切核糖核酸酶,在 AACU 处切割 RNA。纯化的 MazF-lp 的内切核糖核酸酶活性被纯化的 MazE-lp 抑制。我们发现 MazE-lp 和 MazEF-lp 复合物特异性结合到 mazEF-lp 操纵子 5'-非翻译区中存在的回文序列。MazE-lp 和 MazEF-lp 可能都通过特异性结合这些基因 5'-UTR 中的回文序列,作为 mazEF-lp 操纵子和其他基因(包括 icmR)的抑制剂,icmR 的基因产物作为 IcmQ 孔形成蛋白的分泌伴侣发挥作用。