Chen Fan, Cocaign-Bousquet Muriel, Girbal Laurence, Nouaille Sébastien
TBI, CNRS, INRAE, INSA, Université de Toulouse, Toulouse, France.
Front Microbiol. 2022 Dec 21;13:1088941. doi: 10.3389/fmicb.2022.1088941. eCollection 2022.
A set of 41 synthetic 5'UTRs with different theoretical translation initiation rates were generated to explore the role of 5'UTRs in the regulation of protein levels in . The roles of the synthetic 5'UTRs in regulating the expression of different reporter genes were analyzed . Protein levels varied substantially between the different constructs but for most of the 5'UTRs, protein levels were not correlated with theoretical translation initiation rates. Large variations in mRNA concentrations were measured with the different 5'UTRs even though the same concentration of transcription inducer was used in each case. 5'UTRs were also found to strongly affect mRNA stability, and these changes in mRNA stability often contributed to observed differences in mRNA concentration. Unexpectedly, the effect of the 5'UTRs on mRNA half-lives was found to vary depending on the downstream reporter gene. These results clearly demonstrate that 5'UTRs contribute to gene expression regulation at the level of translation initiation and of mRNA stability, to an extent that depends on the nature of the downstream gene.
生成了一组41个具有不同理论翻译起始率的合成5'非翻译区（5'UTR），以探究5'UTR在调控[具体物种]蛋白质水平中的作用。分析了合成5'UTR在调控不同报告基因表达中的作用。不同构建体之间的蛋白质水平差异很大，但对于大多数5'UTR而言，蛋白质水平与理论翻译起始率无关。尽管每种情况下使用的转录诱导剂浓度相同，但用不同的5'UTR测量到的mRNA浓度存在很大差异。还发现5'UTR对mRNA稳定性有强烈影响，而mRNA稳定性的这些变化常常导致观察到的mRNA浓度差异。出乎意料的是，发现5'UTR对mRNA半衰期的影响因下游报告基因而异。这些结果清楚地表明，5'UTR在翻译起始和mRNA稳定性水平上对基因表达调控有贡献，其程度取决于下游基因的性质。
