Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 117997 Moscow, Russia.
Int J Mol Sci. 2021 Sep 7;22(18):9679. doi: 10.3390/ijms22189679.
The autogenous regulation of ribosomal protein (r-protein) synthesis plays a key role in maintaining the stoichiometry of ribosomal components in bacteria. In this work, taking the gene as a classic example, we addressed for the first time the in vivo regulation of r-protein synthesis in the mycobacteria () and (). We used a strategy based on chromosomally integrated reporters under the control of the regulatory regions and the ectopic expression of S15 to measure its impact on the reporter expression. Because the use of as a host appeared inefficient, a fluorescent reporter system was developed by inserting or fusions into the chromosome and expressing S15 or S15 in trans from a novel replicative shuttle vector, pAMYC. The results of the eGFP expression measurements in cells provided evidence that the gene in and was feedback-regulated at the translation level. The mutagenic analysis showed that the folding of 5'UTR in a pseudoknot appeared crucial for repression by both S15 and S15, thus indicating a striking resemblance of the feedback control in mycobacteria and in .
核糖体蛋白 (r-protein) 合成的自体调节在维持细菌核糖体成分的化学计量比方面起着关键作用。在这项工作中，我们以经典的 基因为例，首次解决了分枝杆菌（）和（）中 r 蛋白合成的体内调节问题。我们使用基于整合在染色体上的报告基因的策略，这些报告基因受 调控区的控制，并异位表达 S15，以测量其对报告基因表达的影响。由于使用 作为宿主似乎效率低下，因此通过将 或 融合插入 染色体，并从新型复制穿梭载体 pAMYC 中外源表达 S15 或 S15，开发了一个荧光报告系统。在 细胞中进行 eGFP 表达测量的结果提供了证据，表明 和 中的 基因在翻译水平受到反馈调节。突变分析表明， 5'UTR 形成假结的折叠对于 S15 和 S15 的抑制都是至关重要的，因此表明分枝杆菌和 中的 反馈控制具有惊人的相似性。
