Department of Biochemistry, University of Bayreuth, 95447 Bayreuth, Germany.
Bayreuth Center for Biochemistry & Molecular Biology, Universität Bayreuth, 95447 Bayreuth, Germany.
Nucleic Acids Res. 2024 Sep 9;52(16):10017-10028. doi: 10.1093/nar/gkae678.
Vital organismal processes, including development, differentiation and adaptation, involve altered gene expression. Although expression is frequently controlled at the transcriptional stage, various regulation mechanisms operate at downstream levels. Here, we leverage the photoreceptor NmPAL to optogenetically induce RNA refolding and the translation of bacterial mRNAs. Blue-light-triggered NmPAL binding disrupts a cis-repressed mRNA state, thereby relieves obstruction of translation initiation, and upregulates gene expression. Iterative probing and optimization of the circuit, dubbed riboptoregulator, enhanced induction to 30-fold. Given action at the mRNA level, the riboptoregulator can differentially regulate individual structural genes within polycistronic operons. Moreover, it is orthogonal to and can be wed with other gene-regulatory circuits for nuanced and more stringent gene-expression control. We thus advance the pAurora2 circuit that combines transcriptional and translational mechanisms to optogenetically increase bacterial gene expression by >1000-fold. The riboptoregulator strategy stands to upgrade numerous regulatory circuits and widely applies to expression control in microbial biotechnology, synthetic biology and materials science.
生命有机过程，包括发育、分化和适应，都涉及基因表达的改变。尽管表达通常在转录水平上受到控制，但各种调节机制在下游水平起作用。在这里，我们利用光受体 NmPAL 来光遗传学诱导 RNA 重折叠和细菌 mRNA 的翻译。蓝光触发的 NmPAL 结合破坏了顺式抑制的 mRNA 状态，从而解除了翻译起始的阻碍，并上调了基因表达。对该电路（称为核糖调控器）进行了迭代探测和优化，将诱导倍数提高到 30 倍。由于作用于 mRNA 水平，核糖调控器可以在多顺反子操纵子内差异调节单个结构基因。此外，它与其他基因调控回路正交，可以与之结合，用于更精细和更严格的基因表达控制。因此，我们改进了 pAurora2 电路，该电路结合了转录和翻译机制，通过光遗传学将细菌基因表达提高了 1000 多倍。核糖调控器策略有望升级许多调节回路，并广泛应用于微生物生物技术、合成生物学和材料科学中的表达控制。
