Modular and Single-Cell Sensors of Bacterial Ser/Thr Kinase Activity,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Modular and Single-Cell Sensors of Bacterial Ser/Thr Kinase Activity
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2021-08-31 , DOI: 10.1021/acssynbio.1c00250
Christine R Zheng _{1,

2} , Abhyudai Singh ₃ , Alexandra Libby ₄ , Pamela A Silver _{1,

2} , Elizabeth A Libby _{1,

2}

Affiliation

At the single-cell level, protein kinase activity is typically inferred from downstream transcriptional reporters. However, promoters are often coregulated by several pathways, making the activity of a specific kinase difficult to deconvolve. Here, we present modular, direct, and specific sensors of bacterial kinase activity, including FRET-based sensors, as well as a synthetic transcription factor based on the lactose repressor (LacI) that has been engineered to respond to phosphorylation. We demonstrate the utility of these sensors in measuring the activity of PrkC, a conserved bacterial Ser/Thr kinase, in different growth conditions from single cells to colonies. We also show that PrkC activity increases in response to a cell-wall active antibiotic that blocks the late steps in peptidoglycan synthesis (cefotaxime), but not the early steps (fosfomycin). These sensors have a modular design that should generalize to other bacterial signaling systems in the future.

中文翻译：

细菌丝氨酸/苏氨酸激酶活性的模块化和单细胞传感器

在单细胞水平上，蛋白激酶活性通常是从下游转录报告基因中推断出来的。然而，启动子通常由几种途径共同调节，使得特定激酶的活性难以去卷积。在这里，我们展示了细菌激酶活性的模块化、直接和特异性传感器，包括基于 FRET 的传感器，以及基于乳糖阻遏物 (LacI) 的合成转录因子，该转录因子经过工程改造以响应磷酸化。我们展示了这些传感器在测量 PrkC（一种保守的细菌 Ser/Thr 激酶）在从单细胞到菌落的不同生长条件下的活性方面的实用性。我们还表明，PrkC 活性随着细胞壁活性抗生素的增加而增加，这种抗生素阻断了肽聚糖合成的后期步骤（头孢噻肟），但不是早期步骤（磷霉素）。这些传感器具有模块化设计，将来应该可以推广到其他细菌信号系统。

更新日期：2021-09-17

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11