Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA; Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
J Mol Biol. 2019 Feb 1;431(3):615-624. doi: 10.1016/j.jmb.2018.11.004. Epub 2018 Nov 8.
RNase P catalyzes removal of the 5' leader from precursor tRNAs (pre-tRNAs) in all three domains of life. Some eukaryotic cells contain multiple forms of the protein-only RNase P (PRORP) variant, prompting efforts to unravel this seeming redundancy. Previous studies concluded that there were only modest differences in the processing of typical pre-tRNAs by the three isoforms in Arabidopsis thaliana [AtPRORP1 (organellar), AtPRORP2 and AtPRORP3 (nuclear)]. Here, we investigated if different physical attributes of the three isoforms might engender payoffs under specific conditions. Our temperature-activity profiling studies revealed that AtPRORPs display substrate-identity dependent behavior at elevated temperatures (37-45 °C), with the organellar variant outperforming the nuclear counterparts. Echoing these findings, molecular dynamics simulations revealed that AtPRORP2 relative to AtPRORP1 samples a wider conformational ensemble that deviates from the crystal structure. Results from our biochemical studies and molecular dynamics simulations support the idea that AtPRORPs have overlapping but not necessarily redundant attributes and inspire new perspectives on the suitability of each variant to perform its function(s) in a specific cellular locale.
RNase P 催化所有三个生命领域中前体 tRNA(pre-tRNA)的 5' 引导序列的去除。一些真核细胞含有多种蛋白质-only RNase P(PRORP)变体,这促使人们努力揭开这种看似冗余的现象。以前的研究得出结论,拟南芥中三种同工酶在典型 pre-tRNA 的加工中只有适度的差异 [AtPRORP1(细胞器)、AtPRORP2 和 AtPRORP3(核)]。在这里,我们研究了三种同工酶的不同物理属性是否可能在特定条件下产生收益。我们的温度活性谱研究表明,AtPRORPs 在高温(37-45°C)下表现出底物依赖性行为,细胞器变体的表现优于核变体。这些发现与分子动力学模拟结果相呼应,表明 AtPRORP2 相对于 AtPRORP1 会采样更广泛的构象集合,从而偏离晶体结构。我们的生化研究和分子动力学模拟结果支持这样一种观点,即 AtPRORPs 具有重叠但不一定冗余的属性,并为每个变体在特定细胞位置执行其功能的适宜性提供了新的视角。