The signal processing capabilities of bacterial signaling networks offer immense potential for advanced phospho-signaling systems for synthetic biology. Emerging models suggest that complex development may require interconnections between what were once thought to be isolated signaling arrays. For example, Caulobacter crescentus achieves the feat of asymmetric division by utilizing a novel pseudokinase DivL, which senses the output of one signaling pathway to modulate a second pathway. It has been proposed that DivL reverses signal flow by exploiting conserved kinase conformational changes and protein–protein interactions. We engineered a series of DivL-based modulators to synthetically stimulate reverse signaling of the network in vivo. Stimulation of conformational changes through the DivL signal transmission helix resulted in changes to hallmark features of the network: C. crescentus motility and DivL accumulation at the cell poles. Additionally, mutations to a conserved PAS sensor transmission motif disrupted reverse signaling flow in vivo. We propose that synthetic stimulation and sensor disruption provide strategies to define signaling circuit organization principles for the rational design and validation of synthetic pathways.

中文翻译：

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细菌多激酶网络内信号流的综合控制。

细菌信号网络的信号处理能力为用于合成生物学的高级磷酸信号系统提供了巨大的潜力。新兴模型表明，复杂的开发可能需要将曾经被认为是孤立的信号阵列之间的互连。例如，新月形杆菌通过利用一种新型的假激酶DivL实现了不对称分裂的壮举，该假激酶可检测一个信号通路的输出以调节第二个通路。有人提出DivL通过利用保守的激酶构象变化和蛋白质间相互作用来逆转信号流。我们设计了一系列基于DivL的调节剂，以在体内综合刺激网络的反向信号传递。通过DivL信号传输螺旋刺激构象变化，导致网络标志性特征发生变化：弯月梭菌运动性和DivL在细胞两极的蓄积。另外，保守的PAS传感器传递基序的突变破坏了体内反向信号流。我们建议合成刺激和传感器破坏提供策略来定义信号通路组织原理，以合理设计和验证合成途径。