Raw data for manuscript: A biosensor for direct visualisation of auxin

Raw data ressources for Herud et al, 2021

Summary of the work see the text at the end of this section. For details on imaging Methods see the corresponding publication.

All underlying raw data presented in this work is deposited here except:

1) the FACS data which is sepeartedly uploaded in "FlowRepository.org" under identifier "FR-FCM-Z3FL"

2) the structural data which is deposited with the pdb under indetifiers: 6EJW, 6EJZ, 6ENI, 6EKP, 6ENN, 6ELB, 6ELF, 6ELG

For deposition at zenodo.org 3 archive files have been generated and are accessible here:

1) "Herud_et_al_2021__figures_3_and_4_raw_data__v1.zip"

containing the following folders:

a) "figure_3c-d__DMSO_control"

b) "figure_3c-d__import _IAA_1h"

c) "figure_3e-f__import _AA_10_min"

d) "figure_3g-h__export_10min_pre_wash_out"

e) "figure_3g-h__export_1h_pre_wash_out"

f) "figure_3i-j__ER"

g) "figure_4a-b__BFA"

h) "figure_4c-d__gravitropism"

2) "Herud_et_al_2021__suppl_figures_S1_S3-S5_S7_and_table_S2_raw_data__v1.zip"

containing the following folders:

a) "figure_S1__variants"

b) "figure_S3a-c__FRET"

c) "figure_S3e-j__linker"

d) "figure_S4a__pH"

e) "figure_S4b__agents"

f) "figure_S4c__pH_ITC"

g) "figure_S5__compounds"

h) "figure_S8__PIN"

i) "table_S2__variants_ITC"

3) "Herud_et_al_2021__ImageJ_analysis_macros__v1.zip"

containing the following files:

a) "AuxSen_GUI_v-1-0.ijm"

b) "analysis_ER_201123.ijm"

Work Summary:

In plants, one of the most important regulative small molecules is the auxin indole-3-acetic acid (IAA). Its dynamic redistribution plays an essential role in virtually every aspect of plant life, ranging from cell shape and division to organogenesis and to responses to light and gravity . So far, the spatial and temporal distribution of auxin at cellular resolution could not be determined directly. Instead it has been inferred from the visualisation of irreversible processes involving the endogenous auxin response machinery . This detection system fails to record transient changes. Here we report on a genetically encoded biosensor for quantitative in vivo visualisation of auxin distributions. The sensor is based on the E. coli tryptophan repressor (TrpR) whose binding pocket was engineered to be specific to IAA. The sensor employs FRET as readout after coupled to optimally selected fluorescent proteins. This sensor, unlike previous systems, enables direct monitoring of fast uptake and clearance of auxin by individual cells and within cell compartments in planta, responding to the graded spatial distribution along the root axis and its perturbation by transport inhibitors as well as the rapid and reversible redistribution of endogenous auxin in response to changes in gravity vectors. Thus, our auxin sensor enables real-time monitoring of auxin concentrations at a (sub)cellular resolution and their changes in time and space during a plant ́s entire life.