Artificial Protein-Responsive Riboswitches Upregulate Non-AUG Translation Initiation in Yeast.,ACS Synthetic Biology

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Artificial Protein-Responsive Riboswitches Upregulate Non-AUG Translation Initiation in Yeast.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-06-12 , DOI: 10.1021/acssynbio.0c00206
Fumihiro Horie ₁ , Kei Endo ₁ , Koichi Ito ₁

Affiliation

Artificial control of gene expression is one of the core technologies for engineering biological systems. Riboswitches are cis-acting elements on mRNA that regulate gene expression in a ligand-dependent manner often seen in prokaryotes, but rarely in eukaryotes. Because of the poor variety of such elements available in eukaryotic systems, the number of artificially engineered eukaryotic riboswitches, especially of the upregulation type, is still limited. Here, we developed a design principle for upregulation-type riboswitches that utilize non-AUG initiation induced by ribosomal stalling in a ligand-dependent manner in Saccharomyces cerevisiae. Our design principle simply required the proper positioning of a near-cognate start codon relative to the RNA aptamer. Intriguingly, the CUG codon was the most preferable for non-AUG ON switches in terms of output level and switch performance. This work establishes novel choices for artificial genetic control in eukaryotes with versatile potential for industrial and biomedical applications as well as basic research.

中文翻译：

人工蛋白响应性核糖开关上调酵母中的非 AUG 翻译起始。

基因表达的人工控制是工程生物系统的核心技术之一。核糖开关是 mRNA 上的顺式作用元件，以配体依赖性方式调节基因表达，这种方式常见于原核生物，但在真核生物中很少见。由于真核系统中可用的此类元件种类很少，人工改造的真核核糖开关，尤其是上调类型的数量仍然有限。在这里，我们开发了上调型核糖开关的设计原理，该原理利用酿酒酵母中核糖体停滞以配体依赖性方式诱导的非AUG起始。我们的设计原则只是要求近同源起始密码子相对于 RNA 适体的正确定位。有趣的是，就输出水平和开关性能而言，CUG 密码子对于非 AUG ON 开关来说是最优选的。这项工作为真核生物人工遗传控制建立了新的选择，具有工业和生物医学应用以及基础研究的多种潜力。

更新日期：2020-07-17

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