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Artificial signaling in mammalian cells enabled by prokaryotic two-component system.
Nature Chemical Biology ( IF 14.8 ) Pub Date : 2019-12-16 , DOI: 10.1038/s41589-019-0429-9
Alain Mazé 1 , Yaakov Benenson 1
Affiliation  

Augmenting live cells with new signal transduction capabilities is a key objective in genetic engineering and synthetic biology. We showed earlier that two-component signaling pathways could function in mammalian cells, albeit while losing their ligand sensitivity. Here, we show how to transduce small-molecule ligands in a dose-dependent fashion into gene expression in mammalian cells using two-component signaling machinery. First, we engineer mutually complementing truncated mutants of a histidine kinase unable to dimerize and phosphorylate the response regulator. Next, we fuse these mutants to protein domains capable of ligand-induced dimerization, which restores the phosphoryl transfer in a ligand-dependent manner. Cytoplasmic ligands are transduced by facilitating mutant dimerization in the cytoplasm, while extracellular ligands trigger dimerization at the inner side of a plasma membrane. These findings point to the potential of two-component regulatory systems as enabling tools for orthogonal signaling pathways in mammalian cells.

中文翻译:

哺乳动物细胞中的人工信号通过原核两组分系统实现。

利用新的信号转导功能增强活细胞是基因工程和合成生物学的主要目标。我们较早地表明,尽管失去了配体敏感性,但两组分信号通路仍可以在哺乳动物细胞中发挥作用。在这里,我们展示了如何使用两组分信号转导机制以剂量依赖性方式将小分子配体转导到哺乳动物细胞中的基因表达中。首先,我们设计了互为补充的组氨酸激酶的截短突变体,这些突变体不能使响应调节剂二聚化和磷酸化。接下来,我们将这些突变体融合到能够进行配体诱导的二聚化的蛋白质域中,从而以依赖配体的方式恢复磷酸基转移。通过促进细胞质中的突变体二聚化来转导细胞质配体,而细胞外配体则触发质膜内侧的二聚化。这些发现表明,两组分调节系统作为哺乳动物细胞中正交信号通路的使能工具的潜力。
更新日期:2019-12-17
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