Institute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, Germany.
Bioinformatics Group, Department of Computer Science and Interdisciplinary Center for Bioinformatics, Leipzig University, Härtelstr. 16-18, 04107 Leipzig, Germany.
Nucleic Acids Res. 2021 Feb 22;49(3):1784-1800. doi: 10.1093/nar/gkaa1282.
We describe a synthetic riboswitch element that implements a regulatory principle which directly addresses an essential tRNA maturation step. Constructed using a rational in silico design approach, this riboswitch regulates RNase P-catalyzed tRNA 5'-processing by either sequestering or exposing the single-stranded 5'-leader region of the tRNA precursor in response to a ligand. A single base pair in the 5'-leader defines the regulatory potential of the riboswitch both in vitro and in vivo. Our data provide proof for prior postulates on the importance of the structure of the leader region for tRNA maturation. We demonstrate that computational predictions of ligand-dependent structural rearrangements can address individual maturation steps of stable non-coding RNAs, thus making them amenable as promising target for regulatory devices that can be used as functional building blocks in synthetic biology.
我们描述了一个合成的核糖开关元件，它实现了一个直接针对必需的 tRNA 成熟步骤的调节原理。该核糖开关使用合理的计算设计方法构建，通过响应配体，将 tRNA 前体的单链 5'-leader 区域隔离或暴露出来，从而调节 RNase P 催化的 tRNA 5'加工。5'-leader 中的一个碱基对在体外和体内都定义了核糖开关的调节潜力。我们的数据为关于 leader 区域结构对 tRNA 成熟重要性的先前假设提供了证据。我们证明，对配体依赖性结构重排的计算预测可以解决稳定非编码 RNA 的个别成熟步骤，从而使它们成为有希望的调节装置的潜在靶点，这些调节装置可以作为合成生物学中的功能构建块。
