title: A multi-colour/multi-affinity marker set to visualize phosphoinositide dynamics in Arabidopsis.
abstract: Phosphatidylinositolphosphates (PIPs) are phospholipids that contain a phosphorylated inositol head group. PIPs represent a minor fraction of total phospholipids, but are involved in many regulatory processes, such as cell signalling and intracellular trafficking. Membrane compartments are enriched or depleted in specific PIPs, providing a unique composition for these compartments and contributing to their identity. The precise subcellular localization and dynamics of most PIP species is not fully understood in plants. Here, we designed genetically encoded biosensors with distinct relative affinities and expressed them stably in Arabidopsis thaliana. Analysis of this multi-affinity 'PIPline' marker set revealed previously unrecognized localization of various PIPs in root epidermis. Notably, we found that PI(4,5)P2 is able to localize PIP2 -interacting protein domains to the plasma membrane in non-stressed root epidermal cells. Our analysis further revealed that there is a gradient of PI4P, with the highest concentration at the plasma membrane, intermediate concentration in post-Golgi/endosomal compartments, and the lowest concentration in the Golgi. Finally, we also found a similar gradient of PI3P from high in late endosomes to low in the tonoplast. Our library extends the range of available PIP biosensors, and will allow rapid progress in our understanding of PIP dynamics in plants.
authors.affiliation: Centre National de la Recherche Scientifique, Institut National de la Recherche Agronomique, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Laboratoire de Reproduction et Développement des Plantes, Université de Lyon, 46 Allée d'Italie, 69364, Lyon Cedex 07, France.
authors.forename: Mathilde Laetitia Audrey
authors.initials: ML
authors.lastname: Simon
authors.forename: Matthieu Pierre
authors.initials: MP
authors.lastname: Platre
authors.forename: Sonia
authors.initials: S
authors.lastname: Assil
authors.forename: Ringo
authors.initials: R
authors.lastname: van Wijk
authors.forename: William Yawei
authors.initials: WY
authors.lastname: Chen
authors.forename: Joanne
authors.initials: J
authors.lastname: Chory
authors.forename: Marlène
authors.initials: M
authors.lastname: Dreux
authors.forename: Teun
authors.initials: T
authors.lastname: Munnik
authors.forename: Yvon
authors.initials: Y
authors.lastname: Jaillais
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism*; D015374:Biosensing Techniques; D002451:Cell Compartmentation; D010716:Phosphatidylinositols / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D010716:Phosphatidylinositols
keywords: Arabidopsis thaliana; endosome; lipid binding domain; lipid signalling; membrane trafficking; phosphoinositide; sensor; technical advance