An orthogonalized PYR1-based CID module with reprogrammable ligand-binding specificity

Abstract

Plants sense abscisic acid (ABA) using chemical-induced dimerization (CID) modules, including the receptor PYR1 and HAB1, a phosphatase inhibited by ligand-activated PYR1. This system is unique because of the relative ease with which ligand recognition can be reprogrammed. To expand the PYR1 system, we designed an orthogonal ‘*’ module, which harbors a dimer interface salt bridge; X-ray crystallographic, biochemical and in vivo analyses confirm its orthogonality. We used this module to create PYR1*MANDI/HAB1* and PYR1*AZIN/HAB1*, which possess nanomolar sensitivities to their activating ligands mandipropamid and azinphos-ethyl. Experiments in Arabidopsis thaliana and Saccharomyces cerevisiae demonstrate the sensitive detection of banned organophosphate contaminants using living biosensors and the construction of multi-input/output genetic circuits. Our new modules enable ligand-programmable multi-channel CID systems for plant and eukaryotic synthetic biology that can empower new plant-based and microbe-based sensing modalities.

Document Details

Document Type
Pub Defense Publication
Publication Date
Oct 23, 2023
Source ID
10.1038/s41589-023-01447-7

Entities

People

  • Aditya S. Vaidya
  • Angélica V. Medina-cucurella
  • Brian F Volkman
  • Dmitri A. Nusinow
  • Francis C. Peterson
  • Ian R Wheeldon
  • Jesús Beltrán
  • Jingde Qiu
  • Sang-Youl Park
  • Sean Cutler
  • Shuang Wei
  • Timothy A Whitehead
  • Zenan Xing

Organizations

  • National Institute on Drug Abuse
  • National Science Foundation
  • United States Department of Defense

Tags

Readers

  • Computer Science/Computer Engineering/Data Science/Digital Signal Processing.
  • Molecular Genetics
  • Molecular and Cellular Biochemistry

Technology Areas

  • Biotechnology
  • Biotechnology - Cancer Biotech