The sequences of the 5′ untranslated regions (5′-UTRs) of mRNA alter gene expression across domains of life. Transcriptional modulators can be easily assayed through transcription termination, but translational regulators often require indirect, laborious methods. We have leveraged RelE’s ribosome-dependent endonuclease activity to develop a quantitative assay to monitor translation initiation of cis-regulatory mRNAs. RelE cleavage accurately reports ligand-dependent changes in ribosome association for two translational riboswitches and provides quantitative information about each switch's sensitivity and range of response. RelE accurately reads out sequence-driven changes in riboswitch specificity and function and is quantitatively dependent upon ligand concentration. RelE cleavage similarly captures differences in translation initiation between yeast 5′-UTR isoforms. RelE cleavage can thus reveal a plethora of information about translation initiation in different domains of life.

中文翻译：

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RelE 裂解对翻译起始的有效定量监测

mRNA 的 5' 非翻译区 (5'-UTR) 的序列改变了生命各个领域的基因表达。转录调节剂可以通过转录终止轻松测定，但翻译调节剂通常需要间接、费力的方法。我们利用 RelE 的核糖体依赖性核酸内切酶活性开发了一种定量检测方法来监测顺式调节 mRNA 的翻译起始。RelE 切割准确地报告了两个翻译核糖开关的核糖体结合的配体依赖性变化，并提供了有关每个开关的灵敏度和响应范围的定量信息。RelE 准确读出核糖开关特异性和功能的序列驱动变化，并在数量上取决于配体浓度。RelE 切割同样捕获酵母 5'-UTR 亚型之间翻译起始的差异。因此，RelE 切割可以揭示有关生命不同领域中翻译起始的大量信息。