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鉴定嗜肺军团菌中的 MazF 同源物,该同源物能在 AACU 序列处切割 RNA。

Identification of MazF Homologue in Legionella pneumophila Which Cleaves RNA at the AACU Sequence.

机构信息

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.

出版信息

J Mol Microbiol Biotechnol. 2018;28(6):269-280. doi: 10.1159/000497146. Epub 2019 Mar 20.

Abstract

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.

摘要

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。

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