Scholarlink

<p>C-di-GMP is a widespread second messenger that coordinates transitions between different lifestyles in bacteria. Levels of c-di-GMP are controlled by complex regulatory networks, and they can vary dynamically over a wide range of concentrations. To enable studies of c-di-GMP regulation under a variety of conditions, here we construct and characterize a large set of FRET-based c-di-GMP biosensors that undergo large FRET signal changes and display a stepwise coverage of diverse binding affinities, thus capable of sensitively detecting diverse cellular c-di-GMP concentrations. We subsequently apply different-affinity FRET biosensors from this toolbox to systematically investigate genome-wide network of c-di-GMP regulation in planktonic <i>Escherichia coli</i> cells by establishing FRET-To-Sort, which relies on FRET-based cell sorting of a barcoded transposon library. We observe prominent enrichment of mutations in two classes of flagellar genes among those affecting c-di-GMP levels, and demonstrate that inhibited flagellar rotation reduces biosynthesis of c-di-GMP due to increased proton motive force.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Toolbox of FRET-based c-di-GMP biosensors and its FRET-To-Sort application for genome-wide mapping of c-di-GMP regulation

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年3月26日

查看全文
  • Liyun Wang,
  • Gabriele Malengo,
  • Ananda Sanches-Medeiros,
  • Xuanlin Chen,
  • Julian Pietsch,
  • Nataliya Teteneva,
  • Silvia González Sierra,
  • Ming C. Hammond,
  • Victor Sourjik

摘要

C-di-GMP is a widespread second messenger that coordinates transitions between different lifestyles in bacteria. Levels of c-di-GMP are controlled by complex regulatory networks, and they can vary dynamically over a wide range of concentrations. To enable studies of c-di-GMP regulation under a variety of conditions, here we construct and characterize a large set of FRET-based c-di-GMP biosensors that undergo large FRET signal changes and display a stepwise coverage of diverse binding affinities, thus capable of sensitively detecting diverse cellular c-di-GMP concentrations. We subsequently apply different-affinity FRET biosensors from this toolbox to systematically investigate genome-wide network of c-di-GMP regulation in planktonic Escherichia coli cells by establishing FRET-To-Sort, which relies on FRET-based cell sorting of a barcoded transposon library. We observe prominent enrichment of mutations in two classes of flagellar genes among those affecting c-di-GMP levels, and demonstrate that inhibited flagellar rotation reduces biosynthesis of c-di-GMP due to increased proton motive force.