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  • Click chemistry-based tracking reveals putative cell wall-located auxin binding sites in expanding cells.

Click chemistry-based tracking reveals putative cell wall-located auxin binding sites in expanding cells.

Scientific reports (2017-11-24)
Jozef Mravec, Stjepan K Kračun, Elena Zemlyanskaya, Maja G Rydahl, Xiaoyuan Guo, Martina Pičmanová, Kasper K Sørensen, Kamil Růžička, William G T Willats
ABSTRACT

Auxin is a key plant regulatory molecule, which acts upon a plethora of cellular processes, including those related to cell differentiation and elongation. Despite the stunning progress in all disciplines of auxin research, the mechanisms of auxin-mediated rapid promotion of cell expansion and underlying rearrangement of cell wall components are poorly understood. This is partly due to the limitations of current methodologies for probing auxin. Here we describe a click chemistry-based approach, using an azido derivative of indole-3-propionic acid. This compound is as an active auxin analogue, which can be tagged in situ. Using this new tool, we demonstrate the existence of putative auxin binding sites in the cell walls of expanding/elongating cells. These binding sites are of protein nature but are distinct from those provided by the extensively studied AUXIN BINDING PROTEIN 1 (ABP1). Using immunohistochemistry, we have shown the apoplastic presence of endogenous auxin epitopes recognised by an anti-IAA antibody. Our results are intriguingly in line with previous observations suggesting some transcription-independent (non-genomic) activity of auxin in cell elongation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Streptavidin−Alkaline Phosphatase from Streptomyces avidinii, lyophilized powder, ≥700 DEA units/mg protein
Sigma-Aldrich
Cy3-alkyne, for copper catalyzed click labeling
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide, ≥97.0% (T)