Arabidopsis Rab-E GTPases exhibit a novel interaction with a plasma-membrane phosphatidylinositol-4-phosphate 5-kinase

Rab GTPases of the Arabidopsis Rab-E subclass are related to mammalian Rab8 and are implicated in membrane trafficking from the Golgi to the plasma membrane. Using a yeast two-hybrid assay, Arabidopsis phosphatidylinositol-4-phosphate 5-kinase 2 (PtdIns(4)P 5-kinase 2; also known as PIP5K2), was shown to interact with all five members of the Rab-E subclass but not with other Rab subclasses residing at the Golgi or trans-Golgi network. Interactions in yeast and in vitro were strongest with RAB-E1d[Q74L] and weakest with the RAB-E1d[S29N] suggesting that PIP5K2 interacts with the GTP-bound form. PIP5K2 exhibited kinase activity towards phosphatidylinositol phosphates with a free 5-hydroxyl group, consistent with PtdIns(4)P 5-kinase activity and this activity was stimulated by Rab binding. Rab-E proteins interacted with PIP5K2 via its membrane occupancy and recognition nexus (MORN) domain which is missing from animal and fungal PtdIns(4)P 5-kinases. In plant cells, GFP:PIP5K2 accumulated at the plasma membrane and caused YFP:RAB-E1d to relocate there from its usual position at the Golgi. GFP:PIP5K2 was rapidly turned over by proteasomal activity in planta, and overexpression of YFP:PIP5K2 caused pleiotropic growth abnormalities in transgenic Arabidopsis. We propose that plant cells exhibit a novel interaction in which PIP5K2 binds GTP-bound Rab-E proteins, which may stimulate temporally or spatially localized PtdIns(4,5)P2 production at the plasma membrane.

[1]  J. Chory,et al.  Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. , 2009, The Plant journal : for cell and molecular biology.

[2]  I. Stenzel,et al.  Type B Phosphatidylinositol-4-Phosphate 5-Kinases Mediate Arabidopsis and Nicotiana tabacum Pollen Tube Growth by Regulating Apical Pectin Secretion[W] , 2008, The Plant Cell Online.

[3]  I. Moore,et al.  The functions of Rab GTPases in plant membrane traffic. , 2008, Current opinion in plant biology.

[4]  R. Malhó,et al.  Arabidopsis Phosphatidylinositol-4-Monophosphate 5-Kinase 4 Regulates Pollen Tube Growth and Polarity by Modulating Membrane Recycling[W] , 2008, The Plant Cell Online.

[5]  M. Okabe,et al.  Meichroacidin Containing the Membrane Occupation and Recognition Nexus Motif Is Essential for Spermatozoa Morphogenesis* , 2008, Journal of Biological Chemistry.

[6]  T. Munnik,et al.  The Arabidopsis Phosphatidylinositol Phosphate 5-Kinase PIP5K3 Is a Key Regulator of Root Hair Tip Growth[W][OA] , 2008, The Plant Cell Online.

[7]  C. Chow,et al.  Rab-A2 and Rab-A3 GTPases Define a trans-Golgi Endosomal Membrane Domain in Arabidopsis That Contributes Substantially to the Cell Plate[W] , 2008, The Plant Cell Online.

[8]  B. Hause,et al.  The Type B Phosphatidylinositol-4-Phosphate 5-Kinase 3 Is Essential for Root Hair Formation in Arabidopsis thaliana[W] , 2008, The Plant Cell Online.

[9]  T. Gadella,et al.  Visualization of phosphatidylinositol 4,5-bisphosphate in the plasma membrane of suspension-cultured tobacco BY-2 cells and whole Arabidopsis seedlings. , 2007, The Plant journal : for cell and molecular biology.

[10]  Byeong Wook Jeon,et al.  Phosphatidylinositol 4,5-bisphosphate is important for stomatal opening. , 2007, The Plant journal : for cell and molecular biology.

[11]  I. Moore,et al.  An ARF-GEF acting at the Golgi and in selective endocytosis in polarized plant cells , 2007, Nature.

[12]  J. Shetty,et al.  Radial spoke protein 44 (human meichroacidin) is an axonemal alloantigen of sperm and cilia. , 2007, Gene.

[13]  Guillermo Ayala,et al.  Loss of endocytic clathrin-coated pits upon acute depletion of phosphatidylinositol 4,5-bisphosphate , 2007, Proceedings of the National Academy of Sciences.

[14]  Imara Y. Perera,et al.  The N-terminal Membrane Occupation and Recognition Nexus Domain of Arabidopsis Phosphatidylinositol Phosphate Kinase 1 Regulates Enzyme Activity* , 2007, Journal of Biological Chemistry.

[15]  K. Mecklenburg.  Drosophila retinophilin contains MORN repeats and is conserved in humans , 2007, Molecular Genetics and Genomics.

[16]  H. Xue,et al.  PIP5K9, an Arabidopsis Phosphatidylinositol Monophosphate Kinase, Interacts with a Cytosolic Invertase to Negatively Regulate Sugar-Mediated Root Growth[W] , 2007, The Plant Cell Online.

[17]  M. Fricker,et al.  Ratiometric Fluorescence‐Imaging Assays of Plant Membrane Traffic Using Polyproteins , 2006, Traffic.

[18]  Peter Novick,et al.  Rabs and their effectors: Achieving specificity in membrane traffic , 2006, Proceedings of the National Academy of Sciences.

[19]  B. Striepen,et al.  A MORN-repeat protein is a dynamic component of the Toxoplasma gondii cell division apparatus , 2006, Journal of Cell Science.

[20]  M. Preuss,et al.  A role for the RabA4b effector protein PI-4Kβ1 in polarized expansion of root hair cells in Arabidopsis thaliana , 2006, The Journal of cell biology.

[21]  C. Pikaard,et al.  Gateway-compatible vectors for plant functional genomics and proteomics. , 2006, The Plant journal : for cell and molecular biology.

[22]  S. Munro,et al.  Organelle identity and the signposts for membrane traffic , 2005, Nature.

[23]  P. De Camilli,et al.  An enzymatic cascade of Rab5 effectors regulates phosphoinositide turnover in the endocytic pathway , 2005, The Journal of cell biology.

[24]  P. Hussey,et al.  A Rab-E GTPase Mutant Acts Downstream of the Rab-D Subclass in Biosynthetic Membrane Traffic to the Plasma Membrane in Tobacco Leaf Epidermisw⃞ , 2005, The Plant Cell Online.

[25]  V. Misra,et al.  Identification and characterization of AtCASP, a plant transmembrane Golgi matrix protein , 2005, Plant Molecular Biology.

[26]  A. Parsons,et al.  Synthetic genetic array analysis of the PtdIns 4-kinase Pik1p identifies components in a Golgi-specific Ypt31/rab-GTPase signaling pathway. , 2004, Molecular biology of the cell.

[27]  Lawrence M. Lifshitz,et al.  Phosphatidylinositol-4,5-Bisphosphate-Rich Plasma Membrane Patches Organize Active Zones of Endocytosis and Ruffling in Cultured Adipocytes , 2004, Molecular and Cellular Biology.

[28]  H. Ohta,et al.  ARC3, a chloroplast division factor, is a chimera of prokaryotic FtsZ and part of eukaryotic phosphatidylinositol-4-phosphate 5-kinase. , 2004, Plant & cell physiology.

[29]  H. Batoko,et al.  Targeting of a Nicotiana plumbaginifolia H+-ATPase to the Plasma Membrane Is Not by Default and Requires Cytosolic Structural Determinants , 2004, The Plant Cell Online.

[30]  P. Hussey,et al.  Arabidopsis homologues of the autophagy protein Atg8 are a novel family of microtubule binding proteins , 2004, FEBS letters.

[31]  S. Munro.  Organelle identity and the organization of membrane traffic , 2004, Nature Cell Biology.

[32]  M. Matteis,et al.  PI-loting membrane traffic , 2004, Nature Cell Biology.

[33]  W. Zwart,et al.  Phosphatidylinositol 4-kinasebeta is critical for functional association of rab11 with the Golgi complex. , 2004, Molecular biology of the cell.

[34]  C. Hawes,et al.  A GFP-based assay reveals a role for RHD3 in transport between the endoplasmic reticulum and Golgi apparatus. , 2004, The Plant journal : for cell and molecular biology.

[35]  T. Munnik,et al.  Phospholipid-based signaling in plants. , 2003, Annual review of plant biology.

[36]  A. Wandinger-Ness,et al.  Human VPS34 and p150 are Rab7 Interacting Partners , 2003, Traffic.

[37]  Shuh Narumiya,et al.  ALS2, a novel guanine nucleotide exchange factor for the small GTPase Rab5, is implicated in endosomal dynamics. , 2003, Human molecular genetics.

[38]  Zhenbiao Yang,et al.  Analysis of the Small GTPase Gene Superfamily of Arabidopsis1 , 2003, Plant Physiology.

[39]  I. Moore,et al.  The Arabidopsis Rab GTPase family: another enigma variation. , 2002, Current opinion in plant biology.

[40]  B. Mueller‐Roeber,et al.  Inositol Phospholipid Metabolism in Arabidopsis. Characterized and Putative Isoforms of Inositol Phospholipid Kinase and Phosphoinositide-Specific Phospholipase C1 , 2002, Plant Physiology.

[41]  Dirk Inzé,et al.  GATEWAY vectors for Agrobacterium-mediated plant transformation. , 2002, Trends in plant science.

[42]  J. Pereira-Leal,et al.  Evolution of the Rab family of small GTP-binding proteins. , 2001, Journal of molecular biology.

[43]  C. Pical,et al.  AtPIP5K1, an Arabidopsis thaliana phosphatidylinositol phosphate kinase, synthesizes PtdIns(3,4)P(2) and PtdIns(4,5)P(2) in vitro and is inhibited by phosphorylation. , 2001, The Biochemical journal.

[44]  C. Hawes,et al.  A Rab1 GTPase Is Required for Transport between the Endoplasmic Reticulum and Golgi Apparatus and for Normal Golgi Movement in Plants , 2000, Plant Cell.

[45]  M. Zerial,et al.  Rabenosyn-5, a Novel Rab5 Effector, Is Complexed with Hvps45 and Recruited to Endosomes through a Fyve Finger Domain , 2000, The Journal of cell biology.

[46]  A. Bogdanove,et al.  AvrPto-dependent Pto-interacting proteins and AvrPto-interacting proteins in tomato. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[47]  M. Iino,et al.  Junctophilins: a novel family of junctional membrane complex proteins. , 2000, Molecular cell.

[48]  P. Novick,et al.  The yeast phosphatidylinositol-4-OH kinase Pik1 regulates secretion at the Golgi , 1999, Nature Cell Biology.

[49]  J. Thorner,et al.  Direct Involvement of Phosphatidylinositol 4-Phosphate in Secretion in the Yeast Saccharomyces cerevisiae * , 1999, The Journal of Biological Chemistry.

[50]  M. Zerial,et al.  Phosphatidylinositol-3-OH kinases are Rab5 effectors , 1999, Nature Cell Biology.

[51]  C. Burd,et al.  Vac1p coordinates Rab and phosphatidylinositol 3-kinase signaling in Vps45p-dependent vesicle docking/fusion at the endosome , 1999, Current Biology.

[52]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[53]  K. Shinozaki,et al.  A gene encoding phosphatidylinositol-4-phosphate 5-kinase is induced by water stress and abscisic acid in Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[54]  Marino Zerial,et al.  EEA1 links PI(3)K function to Rab5 regulation of endosome fusion , 1998, Nature.

[55]  G. Prestwich,et al.  Type I Phosphatidylinositol-4-phosphate 5-Kinases Synthesize the Novel Lipids Phosphatidylinositol 3,5-Bisphosphate and Phosphatidylinositol 5-Phosphate* , 1998, The Journal of Biological Chemistry.

[56]  S. Desrivières,et al.  MSS4, a Phosphatidylinositol-4-phosphate 5-Kinase Required for Organization of the Actin Cytoskeleton in Saccharomyces cerevisiae * , 1998, The Journal of Biological Chemistry.

[57]  G. Prestwich,et al.  The C2 Domains of Rabphilin3A Specifically Bind Phosphatidylinositol 4,5-Bisphosphate Containing Vesicles in a Ca2+-dependent Manner , 1998, The Journal of Biological Chemistry.

[58]  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.

[59]  N. Walworth,et al.  Hydrolysis of GTP by Sec4 protein plays an important role in vesicular transport and is stimulated by a GTPase-activating protein in Saccharomyces cerevisiae. , 1992, Molecular and cellular biology.

[60]  J. Goedhart,et al.  Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells. , 2009, The Plant journal : for cell and molecular biology.

[61]  D. Luo,et al.  OsPIPK1, a Rice Phosphatidylinositol Monophosphate Kinase, Regulates Rice Heading by Modifying the Expression of Floral Induction Genes , 2004, Plant Molecular Biology.

[62]  K. Mikoshiba,et al.  A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications , 2002, Nature Biotechnology.

[63]  V. Olkkonen,et al.  Role of Rab GTPases in membrane traffic. , 1997, International review of cytology.

[64]  M. Karin,et al.  New EMBO Member’s Review THE EMBO JOURNAL , 2022 .