Tags

Type your tag names separated by a space and hit enter

Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells.
Plant J. 2009 Jan; 57(2):356-72.PJ

Abstract

Polyphosphoinositides represent a minor group of phospholipids, accounting for less than 1% of the total. Despite their low abundance, these molecules have been implicated in various signalling and membrane trafficking events. Phosphatidylinositol 4-phosphate (PtdIns4P) is the most abundant polyphosphoinositide. (32)Pi-labelling studies have shown that the turnover of PtdIns4P is rapid, but little is known about where in the cell or plant this occurs. Here, we describe the use of a lipid biosensor that monitors PtdIns4P dynamics in living plant cells. The biosensor consists of a fusion between a fluorescent protein and a lipid-binding domain that specifically binds PtdIns4P, i.e. the pleckstrin homology domain of the human protein phosphatidylinositol-4-phosphate adaptor protein-1 (FAPP1). YFP-PH(FAPP1) was expressed in four plant systems: transiently in cowpea protoplasts, and stably in tobacco BY-2 cells, Medicago truncatula roots and Arabidopsis thaliana seedlings. All systems allowed YFP-PH(FAPP1) expression without detrimental effects. Two distinct fluorescence patterns were observed: labelling of motile punctate structures and the plasma membrane. Co-expression studies with organelle markers revealed strong co-labelling with the Golgi marker STtmd-CFP, but not with the endocytic/pre-vacuolar marker GFP-AtRABF2b. Co-expression with the Ptdins3P biosensor YFP-2 x FYVE revealed totally different localization patterns. During cell division, YFP-PH(FAPP1) showed strong labelling of the cell plate, but PtdIns3P was completely absent from the newly formed cell membrane. In root hairs of M. truncatula and A. thaliana, a clear PtdIns4P gradient was apparent in the plasma membrane, with the highest concentration in the tip. This only occurred in growing root hairs, indicating a role for PtdIns4P in tip growth.

Authors+Show Affiliations

Department of Molecular Cytology, Centre for Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18785997

Citation

Vermeer, Joop E M., et al. "Imaging Phosphatidylinositol 4-phosphate Dynamics in Living Plant Cells." The Plant Journal : for Cell and Molecular Biology, vol. 57, no. 2, 2009, pp. 356-72.
Vermeer JE, Thole JM, Goedhart J, et al. Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells. Plant J. 2009;57(2):356-72.
Vermeer, J. E., Thole, J. M., Goedhart, J., Nielsen, E., Munnik, T., & Gadella, T. W. (2009). Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells. The Plant Journal : for Cell and Molecular Biology, 57(2), 356-72. https://doi.org/10.1111/j.1365-313X.2008.03679.x
Vermeer JE, et al. Imaging Phosphatidylinositol 4-phosphate Dynamics in Living Plant Cells. Plant J. 2009;57(2):356-72. PubMed PMID: 18785997.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells. AU - Vermeer,Joop E M, AU - Thole,Julie M, AU - Goedhart,Joachim, AU - Nielsen,Erik, AU - Munnik,Teun, AU - Gadella,Theodorus W J,Jr Y1 - 2008/10/25/ PY - 2008/9/13/pubmed PY - 2009/2/6/medline PY - 2008/9/13/entrez SP - 356 EP - 72 JF - The Plant journal : for cell and molecular biology JO - Plant J. VL - 57 IS - 2 N2 - Polyphosphoinositides represent a minor group of phospholipids, accounting for less than 1% of the total. Despite their low abundance, these molecules have been implicated in various signalling and membrane trafficking events. Phosphatidylinositol 4-phosphate (PtdIns4P) is the most abundant polyphosphoinositide. (32)Pi-labelling studies have shown that the turnover of PtdIns4P is rapid, but little is known about where in the cell or plant this occurs. Here, we describe the use of a lipid biosensor that monitors PtdIns4P dynamics in living plant cells. The biosensor consists of a fusion between a fluorescent protein and a lipid-binding domain that specifically binds PtdIns4P, i.e. the pleckstrin homology domain of the human protein phosphatidylinositol-4-phosphate adaptor protein-1 (FAPP1). YFP-PH(FAPP1) was expressed in four plant systems: transiently in cowpea protoplasts, and stably in tobacco BY-2 cells, Medicago truncatula roots and Arabidopsis thaliana seedlings. All systems allowed YFP-PH(FAPP1) expression without detrimental effects. Two distinct fluorescence patterns were observed: labelling of motile punctate structures and the plasma membrane. Co-expression studies with organelle markers revealed strong co-labelling with the Golgi marker STtmd-CFP, but not with the endocytic/pre-vacuolar marker GFP-AtRABF2b. Co-expression with the Ptdins3P biosensor YFP-2 x FYVE revealed totally different localization patterns. During cell division, YFP-PH(FAPP1) showed strong labelling of the cell plate, but PtdIns3P was completely absent from the newly formed cell membrane. In root hairs of M. truncatula and A. thaliana, a clear PtdIns4P gradient was apparent in the plasma membrane, with the highest concentration in the tip. This only occurred in growing root hairs, indicating a role for PtdIns4P in tip growth. SN - 1365-313X UR - https://www.unboundmedicine.com/medline/citation/18785997/Imaging_phosphatidylinositol_4_phosphate_dynamics_in_living_plant_cells_ L2 - https://doi.org/10.1111/j.1365-313X.2008.03679.x DB - PRIME DP - Unbound Medicine ER -