Zhao Ye, Lu Meihua, Zhang Hui, Hu Jing, Zhou Congli, Xu Qiang, Ul Hussain Shah Amir Miraj, Xu Hong, Wang Liangyan, Hua Yuejin
Key Laboratory of Chinese Ministry of Agriculture for Nuclear-Agricultural Sciences, Institute of Nuclear-Agricultural Sciences, Zhejiang University, China
Key Laboratory of Chinese Ministry of Agriculture for Nuclear-Agricultural Sciences, Institute of Nuclear-Agricultural Sciences, Zhejiang University, China.
Nucleic Acids Res. 2015 Jun 23;43(11):5550-9. doi: 10.1093/nar/gkv444. Epub 2015 May 4.
RNase J is a conserved ribonuclease that belongs to the β-CASP family of nucleases. It possesses both endo- and exo-ribonuclease activities, which play a key role in pre-rRNA maturation and mRNA decay. Here we report high-resolution crystal structures of Deinococcus radiodurans RNase J complexed with RNA or uridine 5'-monophosphate in the presence of manganese ions. Biochemical and structural studies revealed that RNase J uses zinc ions for two-metal-ion catalysis. One residue conserved among RNase J orthologues (motif B) forms specific electrostatic interactions with the scissile phosphate of the RNA that is critical for the catalysis and product stabilization. The additional manganese ion, which is coordinated by conserved residues at the dimer interface, is critical for RNase J dimerization and exonuclease activity. The structures may also shed light on the mechanism of RNase J exo- and endonucleolytic activity switch.
核糖核酸酶J是一种保守的核糖核酸酶，属于β - CASP核酸酶家族。它同时具有内切核糖核酸酶和外切核糖核酸酶活性，在核糖体RNA前体（pre - rRNA）成熟和信使核糖核酸（mRNA）降解过程中发挥关键作用。在此，我们报道了在锰离子存在下，嗜放射栖热菌核糖核酸酶J与RNA或5'-单磷酸尿苷复合的高分辨率晶体结构。生化和结构研究表明，核糖核酸酶J利用锌离子进行双金属离子催化。在核糖核酸酶J直系同源物中保守的一个残基（基序B）与RNA的可切割磷酸基团形成特定的静电相互作用，这对催化作用和产物稳定至关重要。由二聚体界面处保守残基配位的额外锰离子，对核糖核酸酶J的二聚化和外切核酸酶活性至关重要。这些结构也可能有助于揭示核糖核酸酶J外切核酸酶和内切核酸酶活性转换的机制。
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