Analysis of Membrane Proteins Transport from Endosomal Compartments to Vacuoles.
Arabidopsis
/ genetics
Arabidopsis Proteins
/ genetics
Cell Membrane
/ metabolism
DNA, Bacterial
/ pharmacology
Dexamethasone
/ pharmacology
Endosomes
/ metabolism
Green Fluorescent Proteins
/ genetics
Mutation
Plants, Genetically Modified
/ genetics
Protein Transport
Proteolysis
Vesicular Transport Proteins
/ genetics
FM dye
FREE1
Multivesicular body
Plasma membrane protein
Prevacuolar compartment
Vacuolar degradation
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2020
2020
Historique:
entrez:
8
7
2020
pubmed:
8
7
2020
medline:
19
3
2021
Statut:
ppublish
Résumé
Endocytosis and endosomal trafficking to vacuoles play important roles in regulating the homeostasis of plasma membrane (PM) proteins in plant cells. FREE1 (FYVE domain protein required for endosomal sorting 1) is a plant-unique component of the ESCRT (endosomal sorting complex required for transport) machinery. In free1 mutant plants, PIN-FORMED 2 (PIN2)-GFP was found to mislocalize from the PM to the tonoplast. In this chapter, we describe a detailed protocol for studying vacuolar sorting and degradation of PIN2-GFP by using T-DNA insertional mutants, dexamethasone (DEX) inducible RNAi lines, and other tools, including Fei-Mao (FM) dye staining and dark treatment. By using these methods, we illustrate the endosomal trafficking and vacuolar degradation of PIN2-GFP in plants.
Identifiants
pubmed: 32632801
doi: 10.1007/978-1-0716-0767-1_2
doi:
Substances chimiques
Arabidopsis Proteins
0
DNA, Bacterial
0
FREE1 protein, Arabidopsis
0
PIN2 protein, Arabidopsis
0
T-DNA
0
Vesicular Transport Proteins
0
Green Fluorescent Proteins
147336-22-9
Dexamethasone
7S5I7G3JQL
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM