Waldminghaus Torsten, Kortmann Jens, Gesing Stefan, Narberhaus Franz
Lehrstuhl für Biologie der Mikroorganismen, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Biol Chem. 2008 Oct;389(10):1319-26. doi: 10.1515/BC.2008.150.
Structured RNAs with fundamental sensory and regulatory potential have been discovered in all kingdoms of life. Bacterial RNA thermometers are located in the 5'-untranslated region of certain heat shock and virulence genes. They regulate translation by masking the Shine-Dalgarno sequence in a temperature-dependent manner. To engineer RNA-based thermosensors, we used a combination of computer-based rational design and in vivo screening. After only two rounds of selection, several RNA thermometers that are at least as efficient as natural thermometers were obtained. Structure probing experiments revealed temperature-dependent conformational changes in these translational control elements. Our study demonstrates that temperature-controlled RNA elements can be designed by a simple combined computational and experimental approach.
在所有生命王国中都发现了具有基本传感和调节潜力的结构化RNA。细菌RNA温度计位于某些热休克和毒力基因的5'非翻译区。它们通过以温度依赖的方式掩盖Shine-Dalgarno序列来调节翻译。为了构建基于RNA的热传感器,我们结合了基于计算机的理性设计和体内筛选。仅经过两轮筛选,就获得了几种效率至少与天然温度计相当的RNA温度计。结构探测实验揭示了这些翻译控制元件中温度依赖的构象变化。我们的研究表明,可以通过一种简单的计算与实验相结合的方法来设计温度控制的RNA元件。