Kowallis Kimberly A, Duvall Samuel W, Zhao Wei, Childers W Seth
Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
Chevron Science Center, University of Pittsburgh, Pittsburgh, PA, USA.
Methods Mol Biol. 2020;2077:141-163. doi: 10.1007/978-1-4939-9884-5_10.
Two-component systems allow bacteria to respond to changes in environmental or cytosolic conditions through autophosphorylation of a histidine kinase (HK) and subsequent transfer of the phosphate group to its downstream cognate response regulator (RR). The RR then elicits a cellular response, commonly through regulation of transcription. Engineering two-component system signaling networks provides a strategy to study bacterial signaling mechanisms related to bacterial cell survival, symbiosis, and virulence, and to develop sensory devices in synthetic biology. Here we focus on the principles for engineering the HK to identify unknown signal inputs, test signal transmission mechanisms, design small molecule sensors, and rewire two-component signaling networks.
双组分系统使细菌能够通过组氨酸激酶（HK）的自磷酸化以及随后将磷酸基团转移至其下游同源应答调节因子（RR）来响应环境或胞质溶胶条件的变化。然后，RR通常通过转录调控引发细胞反应。构建双组分系统信号网络为研究与细菌细胞存活、共生和毒力相关的细菌信号传导机制以及在合成生物学中开发传感装置提供了一种策略。在这里，我们重点关注构建HK的原理，以识别未知信号输入、测试信号传输机制、设计小分子传感器以及重新连接双组分信号网络。
