Dynamic Turnover of PI4P and PI(4,5)P2 under Hypoxia Controls Electrostatic Plasma Membrane Targeting

Phosphatidylinositol 4-phosphate (PI4P) and phosphatidylinositol 4,5-biphosphate (PIP2) are key phosphoinositides that determine the identity of the plasma membrane (PM) and regulate numerous key biological events there. To date, mechanisms regulating the homeostasis and dynamic turnover of PM PI4P and PIP2 in response to various physiological conditions and stresses remain to be fully elucidated. Here we report that hypoxia in Drosophila induces acute and reversible depletion of PM PI4P and PIP2 that severely disrupts the electrostatic PM targeting of multiple polybasic polarity proteins. Genetically encoded ATP sensors confirmed that hypoxia induces acute and reversible reduction of cellular ATP levels which showed a strong real-time correlation with the levels of PM PI4P and PIP2 in cultured cells. By combining genetic manipulations with quantitative imaging assays we showed that PI4KIIIα, as well as Rbo/EFR3 and TTC7 that are essential for targeting PI4KIIIα to PM, are required for maintaining the homeostasis and dynamic turnover of PM PI4P and PIP2 under normoxia and hypoxia. Our results revealed that in cells challenged by energetic stresses triggered by hypoxia, ATP inhibition and possibly ischemia, dramatic turnover of PM PI4P and PIP2 could have profound impact on many cellular processes including electrostatic PM targeting of numerous polybasic proteins.

[1]  Yang Hong,et al.  Electrostatic plasma membrane targeting contributes to Dlg function in cell polarity and tumorigenesis. , 2021, Development.

[2]  A. Molina.  Mitochondrial Dysfunction , 2021, Encyclopedia of Gerontology and Population Aging.

[3]  P. Raghu,et al.  A PI4KIIIα protein complex is required for cell viability during Drosophila wing development. , 2020, Developmental biology.

[4]  J. Burke,et al.  Novel roles of phosphoinositides in signaling, lipid transport, and disease. , 2020, Current opinion in cell biology.

[5]  Y. Wu,et al.  A polybasic domain in aPKC mediates Par6-dependent control of membrane targeting and kinase activity , 2019, bioRxiv.

[6]  T. Balla,et al.  Defining the subcellular distribution and metabolic channeling of phosphatidylinositol , 2019, bioRxiv.

[7]  Gerald R. V. Hammond,et al.  Probing the subcellular distribution of phosphatidylinositol reveals a surprising lack at the plasma membrane , 2019, bioRxiv.

[8]  T. Balla,et al.  Phosphatidylinositol 4,5‐bisphosphate controls Rab7 and PLEKHM1 membrane cycling during autophagosome–lysosome fusion , 2019, The EMBO journal.

[9]  S. Simon,et al.  Escherichia coli as a platform for the study of phosphoinositide biology , 2019, Science Advances.

[10]  Shao Li,et al.  An efficient and multiple target transgenic RNAi technique with low toxicity in Drosophila , 2018, Nature Communications.

[11]  M. Jaiswal,et al.  Regulation of PI4P levels by PI4KIIIα during G-protein-coupled PLC signaling in Drosophila photoreceptors , 2018, Journal of Cell Science.

[12]  L. Cantley,et al.  PIP4Ks Suppress Insulin Signaling through a Catalytic-Independent Mechanism , 2018, bioRxiv.

[13]  M. Raghunath,et al.  RGB-Color Intensiometric Indicators to Visualize Spatiotemporal Dynamics of ATP in Single Cells. , 2018, Angewandte Chemie.

[14]  Gerald R. V. Hammond,et al.  Genetically encoded lipid biosensors , 2018, Molecular biology of the cell.

[15]  Yang Hong.  aPKC: the Kinase that Phosphorylates Cell Polarity , 2018, F1000Research.

[16]  G. Fairn,et al.  PI(4,5)P2 controls plasma membrane PI4P and PS levels via ORP5/8 recruitment to ER–PM contact sites , 2018, The Journal of cell biology.

[17]  R. Hardie,et al.  Genetic dissection of the phosphoinositide cycle in Drosophila photoreceptors , 2018, Journal of Cell Science.

[18]  P. De Camilli,et al.  Architecture of the human PI4KIIIα lipid kinase complex , 2017, Proceedings of the National Academy of Sciences.

[19]  J. Liou,et al.  RASSF4 controls SOCE and ER–PM junctions through regulation of PI(4,5)P2 , 2017, The Journal of cell biology.

[20]  T. Uemura,et al.  Mitochondrial dysfunction induces dendritic loss via eIF2α phosphorylation , 2017, The Journal of cell biology.

[21]  P. De Camilli,et al.  Endosome-ER Contacts Control Actin Nucleation and Retromer Function through VAP-Dependent Regulation of PI4P , 2016, Cell.

[22]  V. Haucke,et al.  A phosphoinositide conversion mechanism for exit from endosomes , 2016, Nature.

[23]  P. De Camilli,et al.  The leukodystrophy protein FAM126A/Hyccin regulates PI4P synthesis at the plasma membrane , 2015, Nature Cell Biology.

[24]  Palladino,et al.  A conserved polybasic domain mediates plasma membrane targeting of Lgl and its regulation by hypoxia , 2022 .

[25]  K. Prehoda,et al.  Establishment of Par-Polarized Cortical Domains via Phosphoregulated Membrane Motifs. , 2015, Developmental cell.

[26]  P. Raghu,et al.  Phosphoinositide signalling in Drosophila. , 2015, Biochimica et biophysica acta.

[27]  P. De Camilli,et al.  Plasticity of PI4KIIIα interactions at the plasma membrane , 2015, EMBO reports.

[28]  B. Hille,et al.  Golgi and plasma membrane pools of PI(4)P contribute to plasma membrane PI(4,5)P2 and maintenance of KCNQ2/3 ion channel current , 2014, Proceedings of the National Academy of Sciences.

[29]  A. Guichet,et al.  PI(4,5)P2 Produced by the PI4P5K SKTL Controls Apical Size by Tethering PAR-3 in Drosophila Epithelial Cells , 2014, Current Biology.

[30]  T. Balla,et al.  A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi , 2014, The Journal of cell biology.

[31]  P. De Camilli,et al.  Structural insights into assembly and regulation of the plasma membrane phosphatidylinositol 4-kinase complex. , 2014, Developmental cell.

[32]  Masasuke Yoshida,et al.  Evaluation of intramitochondrial ATP levels identifies G0/G1 switch gene 2 as a positive regulator of oxidative phosphorylation , 2013, Proceedings of the National Academy of Sciences.

[33]  T. Balla,et al.  Phosphoinositides: tiny lipids with giant impact on cell regulation. , 2013, Physiological reviews.

[34]  Sang Yoon Lee,et al.  PtdIns4P synthesis by PI4KIIIα at the plasma membrane and its impact on plasma membrane identity , 2012, The Journal of cell biology.

[35]  J. A. Brill,et al.  Type II phosphatidylinositol 4-kinase regulates trafficking of secretory granule proteins in Drosophila , 2012, Development.

[36]  H. Krämer,et al.  Type II phosphatidylinositol 4-kinase regulates trafficking of secretory granule proteins in Drosophila , 2012, Journal of Cell Science.

[37]  T. Balla,et al.  PI4P and PI(4,5)P2 Are Essential But Independent Lipid Determinants of Membrane Identity , 2012, Science.

[38]  Juan Huang,et al.  Differential regulation of adherens junction dynamics during apical–basal polarization , 2011, Journal of Cell Science.

[39]  T. Schüpbach,et al.  Drosophila PI4KIIIalpha is required in follicle cells for oocyte polarization and Hippo signaling , 2011, Development.

[40]  T. Balla,et al.  Dual roles for the Drosophila PI 4-kinase Four wheel drive in localizing Rab11 during cytokinesis , 2009, The Journal of cell biology.

[41]  P. De Camilli,et al.  An electrostatic switch displaces phosphatidylinositol phosphate kinases from the membrane during phagocytosis , 2009, The Journal of cell biology.

[42]  Takeharu Nagai,et al.  Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators , 2009, Proceedings of the National Academy of Sciences.

[43]  Wei Dong,et al.  Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering , 2009, Proceedings of the National Academy of Sciences.

[44]  K. Broadie,et al.  Rolling blackout is required for bulk endocytosis in non-neuronal cells and neuronal synapses , 2009, Journal of Cell Science.

[45]  K. Broadie,et al.  Rolling blackout is required for bulk endocytosis in non-neuronal cells and neuronal synapses , 2009, Journal of Cell Science.

[46]  Tony Yeung,et al.  Membrane Phosphatidylserine Regulates Surface Charge and Protein Localization , 2008, Science.

[47]  Kevan M Shokat,et al.  Phosphatidylinositol 4-Kinase IIIβ Regulates the Transport of Ceramide between the Endoplasmic Reticulum and Golgi* , 2006, Journal of Biological Chemistry.

[48]  T. Balla,et al.  Phosphatidylinositol 4-kinases: old enzymes with emerging functions. , 2006, Trends in cell biology.

[49]  D. Murray,et al.  Plasma membrane phosphoinositide organization by protein electrostatics , 2005, Nature.

[50]  K. Broadie,et al.  Rolling blackout, a newly identified PIP2-DAG pathway lipase required for Drosophila phototransduction , 2004, Nature Neuroscience.

[51]  Jean-Christophe Olivo-Marin,et al.  Extraction of spots in biological images using multiscale products , 2002, Pattern Recognit..

[52]  J. A. Brill,et al.  A phospholipid kinase regulates actin organization and intercellular bridge formation during germline cytokinesis. , 2000, Development.

[53]  T. Deuel.  Phosphatidylinositol 4-kinase , 1998 .

[54]  Péter Várnai,et al.  Visualization of Phosphoinositides That Bind Pleckstrin Homology Domains: Calcium- and Agonist-induced Dynamic Changes and Relationship to Myo-[3H]inositol-labeled Phosphoinositide Pools , 1998, The Journal of cell biology.

[55]  T. Schüpbach,et al.  Ectopic activation of torpedo/Egfr, a Drosophila receptor tyrosine kinase, dorsalizes both the eggshell and the embryo. , 1997, Development.

[56]  L. Cantley,et al.  Phosphoinositide kinases. , 1990, Biochemistry.

[57]  J. Putney,et al.  Effects of antimycin A on receptor-activated calcium mobilization and phosphoinositide metabolism in rat parotid gland. , 1983, Molecular pharmacology.