ArticleProceedings of the National Academy of Sciences of the United States of America2013
Single-particle analysis reveals shutoff control of the Arabidopsis ammonium transporter AMT1;3 by clustering and internalization.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.
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42 citing papers in PubMed, 102 citations in OpenAlex.
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- Nitrogen Journey in Plants: From Uptake to Metabolism, Stress Response, and Microbe Interaction.Biomolecules · 2023Review
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- Primary nutrient sensors in plants.iScience · 2022Review
- Membrane nanodomains and transport functions in plant.Plant physiology · 2021Review
- A proposed role for endomembrane trafficking processes in regulating tonoplast content and vacuole dynamics under ammonium stress conditions in Arabidopsis root cells.Plant signaling & behavior · 2021Article
- Single-Molecule Imaging in Living Plant Cells: A Methodological Review.International journal of molecular sciences · 2021Review
- In vivo single-particle tracking of the aquaporin AtPIP2;1 in stomata reveals cell type-specific dynamics.Plant physiology · 2021Article
- Nitrogen Uptake in Plants: The Plasma Membrane Root Transport Systems from a Physiological and Proteomic Perspective.Plants (Basel, Switzerland) · 2021Review
- Single-particle tracking photoactivated localization microscopy of membrane proteins in living plant tissues.Nature protocols · 2021Article
- Function and Regulation of Ammonium Transporters in Plants.International journal of molecular sciences · 2020Review
- The Nanoscale Organization of the Plasma Membrane and Its Importance in Signaling: A Proteolipid Perspective.Plant physiology · 2020Review
- Dynamics and Endocytosis of Flot1 inInternational journal of molecular sciences · 2020Article
- CesA6 and PGIP2 Endocytosis Involves Different Subpopulations of TGN-Related Endosomes.Frontiers in plant science · 2020Article
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Authors and funding
10 authors at 7 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Ammonium is a preferred source of nitrogen for plants but is toxic at high levels. Plant ammonium transporters (AMTs) play an essential role in NH4(+) uptake, but the mechanism by which AMTs are regulated remains unclear. To study how AMTs are regulated in the presence of ammonium, we used variable-angle total internal reflection fluorescence microscopy and fluorescence cross-correlation spectroscopy for single-particle fluorescence imaging of EGFP-tagged AMT1;3 on the plasma membrane of Arabidopsis root cells at various ammonium levels. We demonstrated that AMT1;3-EGFP dynamically appeared and disappeared on the plasma membrane as moving fluorescent spots in low oligomeric states under N-deprived and N-sufficient conditions. Under external high-ammonium stress, however, AMT1;3-EGFPs were found to amass into clusters, which were then internalized into the cytoplasm. A similar phenomenon also occurred in the glutamine synthetase mutant gln1;2 background. Single-particle analysis of AMT1;3-EGFPs in the clathrin heavy chain 2 mutant (chc2 mutant) and Flotllin1 artificial microRNA (Flot1 amiRNA) backgrounds, together with chemical inhibitor treatments, demonstrated that the endocytosis of AMT1;3 clusters induced by high-ammonium stress could occur mainly through clathrin-mediated endocytic pathways, but the contribution of microdomain-associated endocytic pathway cannot be excluded in the internalization. Our results revealed that the clustering and endocytosis of AMT1;3 provides an effective mechanism by which plant cells can avoid accumulation of toxic levels of ammonium by eliminating active AMT1;3 from the plasma membrane.
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