title: Abscisic acid dynamics in roots detected with genetically encoded FRET sensors.
abstract: Cytosolic hormone levels must be tightly controlled at the level of influx, efflux, synthesis, degradation and compartmentation. To determine ABA dynamics at the single cell level, FRET sensors (ABACUS) covering a range ∼0.2-800 µM were engineered using structure-guided design and a high-throughput screening platform. When expressed in yeast, ABACUS1 detected concentrative ABA uptake mediated by the AIT1/NRT1.2 transporter. Arabidopsis roots expressing ABACUS1-2µ (Kd∼2 µM) and ABACUS1-80µ (Kd∼80 µM) respond to perfusion with ABA in a concentration-dependent manner. The properties of the observed ABA accumulation in roots appear incompatible with the activity of known ABA transporters (AIT1, ABCG40). ABACUS reveals effects of external ABA on homeostasis, that is, ABA-triggered induction of ABA degradation, modification, or compartmentation. ABACUS can be used to study ABA responses in mutants and quantitatively monitor ABA translocation and regulation, and identify missing components. The sensor screening platform promises to enable rapid fine-tuning of the ABA sensors and engineering of plant and animal hormone sensors to advance our understanding of hormone signaling. DOI: http://dx.doi.org/10.7554/eLife.01741.001.
authors.affiliation: Department of Plant Biology, Carnegie Institution for Science, Stanford, United States.
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Jones
authors.forename: Jonas Ah
authors.initials: JA
authors.lastname: Danielson
authors.forename: Shruti N
authors.initials: SN
authors.lastname: Manojkumar
authors.forename: Viviane
authors.initials: V
authors.lastname: Lanquar
authors.forename: Guido
authors.initials: G
authors.lastname: Grossmann
authors.forename: Wolf B
authors.initials: WB
authors.lastname: Frommer
mesh_terms: D000040:Abscisic Acid / Q000032:analysis*; D000818:Animals; D017360:Arabidopsis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D006706:Homeostasis; D010937:Plant Growth Regulators / Q000032:analysis*; D018517:Plant Roots / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified
chemical_list: D005456:Fluorescent Dyes; D010937:Plant Growth Regulators; D000040:Abscisic Acid
keywords: Saccharomyces cerevisiae; drought; germination; hormone; root; transport
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