title: Visualization of phosphatidic acid fluctuations in the plasma membrane of living cells.
abstract: We developed genetically-encoded fluorescent sensors based on Förster Resonance Energy Transfer to monitor phosphatidic acid (PA) fluctuations in the plasma membrane using Spo20 as PA-binding motif. Basal PA levels and phospholipase D activity varied in different cell types. In addition, stimuli that activate PA phosphatases, leading to lower PA levels, increased lamellipodia and filopodia formation. Lower PA levels were observed in the leading edge than in the trailing edge of migrating HeLa cells. In MSC80 and OLN93 cells, which are stable cell lines derived from Schwann cells and oligodendrocytes, respectively, a higher ratio of diacylglycerol to PA levels was demonstrated in the membrane processes involved in myelination, compared to the cell body. We propose that the PA sensors reported here are valuable tools to unveil the role of PA in a variety of intracellular signaling pathways.
authors.affiliation: Centro Regional de Investigaciones Biomédicas and Facultad de Medicina de Albacete, Universidad de Castilla-La Mancha, Albacete, Spain.
authors.forename: José P
authors.initials: JP
authors.lastname: Ferraz-Nogueira
authors.affiliation: Centro de Biología Molecular Severo Ochoa and Departamento de Biología Molecular, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: F Javier
authors.initials: FJ
authors.lastname: Díez-Guerra
authors.affiliation: Centro Regional de Investigaciones Biomédicas and Facultad de Medicina de Albacete, Universidad de Castilla-La Mancha, Albacete, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Llopis
mesh_terms: D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D004075:Diglycerides / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008856:Microscopy, Fluorescence; D010712:Phosphatidic Acids / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D050766:Qb-SNARE Proteins / Q000737:chemistry; D050767:Qc-SNARE Proteins / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D059010:Single-Cell Analysis
chemical_list: D004075:Diglycerides; D008081:Liposomes; D010712:Phosphatidic Acids; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C497084:Spo20 protein, S cerevisiae