Abstract
Cell homeostasis is perturbed when dramatic shifts in the external environment cause the physical-chemical properties inside the cell to change. Experimental approaches for dynamically monitoring these intracellular effects are currently lacking. Here, we leverage the environmental sensitivity and structural plasticity of intrinsically disordered protein regions (IDRs) to develop a FRET biosensor capable of monitoring rapid intracellular changes caused by osmotic stress. The biosensor, named SED1, utilizes the Arabidopsis intrinsically disordered AtLEA4-5 protein expressed in plants under water deficit. Computational modeling and in vitro studies reveal that SED1 is highly sensitive to macromolecular crowding. SED1 exhibits large and near-linear osmolarity-dependent changes in FRET inside living bacteria, yeast, plant, and human cells, demonstrating the broad utility of this tool for studying water-associated stress. This study demonstrates the remarkable ability of IDRs to sense the cellular environment across the tree of life and provides a blueprint for their use as environmentally-responsive molecular tools.
© 2021. The Author(s).
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Arabidopsis / genetics
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Arabidopsis / metabolism
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Arabidopsis Proteins / chemistry
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Arabidopsis Proteins / genetics
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Arabidopsis Proteins / metabolism*
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Binding Sites
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Biosensing Techniques*
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Cell Line, Tumor
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Fluorescence Resonance Energy Transfer
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Gene Expression
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Humans
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Intrinsically Disordered Proteins / chemistry
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Intrinsically Disordered Proteins / genetics
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Intrinsically Disordered Proteins / metabolism*
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Kinetics
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Models, Molecular
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Molecular Chaperones / chemistry
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Molecular Chaperones / genetics
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Molecular Chaperones / metabolism*
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Osmolar Concentration
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Osmotic Pressure*
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Osteoblasts / cytology
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Osteoblasts / metabolism
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Protein Binding
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Protein Conformation
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Protein Interaction Domains and Motifs
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Thermodynamics
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Water / metabolism*
Substances
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AT5G06760 protein, Arabidopsis
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Arabidopsis Proteins
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Intrinsically Disordered Proteins
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Molecular Chaperones
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Recombinant Proteins
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Water