The type Ialpha inositol polyphosphate 4-phosphatase generates and terminates phosphoinositide 3-kinase signals on endosomes and the plasma membrane.

Endosomal trafficking is regulated by the recruitment of effector proteins to phosphatidylinositol 3-phosphate [PtdIns(3)P] on early endosomes. At the plasma membrane, phosphatidylinositol-(3,4)-bisphosphate [PtdIns(3,4)P2] binds the pleckstrin homology (PH) domain-containing proteins Akt and TAPP1. Type Ialpha inositol polyphosphate 4-phosphatase (4-phosphatase) dephosphorylates PtdIns(3,4)P2, forming PtdIns(3)P, but its subcellular localization is unknown. We report here in quiescent cells, the 4-phosphatase colocalized with early and recycling endosomes. On growth factor stimulation, 4-phosphatase endosomal localization persisted, but in addition the 4-phosphatase localized at the plasma membrane. Overexpression of the 4-phosphatase in serum-stimulated cells increased cellular PtdIns(3)P levels and prevented wortmannin-induced endosomal dilatation. Furthermore, mouse embryonic fibroblasts from homozygous Weeble mice, which have a mutation in the type I 4-phosphatase, exhibited dilated early endosomes. 4-Phosphatase translocation to the plasma membrane upon growth factor stimulation inhibited the recruitment of the TAPP1 PH domain. The 4-phosphatase contains C2 domains, which bound PtdIns(3,4)P2, and C2-domain-deletion mutants lost PtdIns(3,4)P2 4-phosphatase activity, did not localize to endosomes or inhibit TAPP1 PH domain membrane recruitment. The 4-phosphatase therefore both generates and terminates phosphoinositide 3-kinase signals at distinct subcellular locations.

[1]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[2]  T. Kreis,et al.  Translocation and clustering of endosomes and lysosomes depends on microtubules , 1987, The Journal of cell biology.

[3]  S. Dowler,et al.  Identification of pleckstrin-homology-domain-containing proteins with novel phosphoinositide-binding specificities. , 2000, The Biochemical journal.

[4]  P. Bastiaens,et al.  Binding of a Diphosphotyrosine-containing Peptide That Mimics Activated Platelet-derived Growth Factor Receptor β Induces Oligomerization of Phosphatidylinositol 3-Kinase* , 1998, The Journal of Biological Chemistry.

[5]  M. J. Clague,et al.  Phosphatidylinositol 3-kinase activity is required for early endosome fusion. , 1995, The Biochemical journal.

[6]  P. Majerus,et al.  Phosphoinositide-specific Inositol Polyphosphate 5-Phosphatase IV Inhibits Akt/Protein Kinase B Phosphorylation and Leads to Apoptotic Cell Death* , 2002, The Journal of Biological Chemistry.

[7]  William Arbuthnot Sir Lane,et al.  Identification of an early endosomal protein regulated by phosphatidylinositol 3-kinase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[8]  C. Mitchell,et al.  Inositol Polyphosphate 5‐Phosphatases: Lipid Phosphatases With Flair , 2002, IUBMB life.

[9]  V. Duronio,et al.  TAPP1 and TAPP2 Are Targets of Phosphatidylinositol 3-Kinase Signaling in B Cells: Sustained Plasma Membrane Recruitment Triggered by the B-Cell Antigen Receptor , 2002, Molecular and Cellular Biology.

[10]  P. Vyas,et al.  Inositol polyphosphate 4-phosphatase type I regulates cell growth downstream of transcription factor GATA-1. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Rein Aasland,et al.  Endosomal Localization of the Autoantigen EEA1 Is Mediated by a Zinc-binding FYVE Finger* , 1996, The Journal of Biological Chemistry.

[12]  H. Geuze,et al.  A novel class of clathrin-coated vesicles budding from endosomes , 1996, The Journal of cell biology.

[13]  P. Majerus,et al.  Isolation and characterization of two 3-phosphatases that hydrolyze both phosphatidylinositol 3-phosphate and inositol 1,3-bisphosphate. , 1991, The Journal of biological chemistry.

[14]  B. Deurs,et al.  A Key to Lysosome Biogenesis , 2000 .

[15]  R. Atkins,et al.  The cDNA Cloning and Characterization of Inositol Polyphosphate 4-Phosphatase Type II , 1997, The Journal of Biological Chemistry.

[16]  P. Majerus,et al.  The Isolation and Characterization of cDNA Encoding Human and Rat Brain Inositol Polyphosphate 4-Phosphatase (*) , 1995, The Journal of Biological Chemistry.

[17]  C. Downes,et al.  Detection of novel intracellular agonist responsive pools of phosphatidylinositol 3,4-bisphosphate using the TAPP1 pleckstrin homology domain in immunoelectron microscopy. , 2004, The Biochemical journal.

[18]  E. Chapman,et al.  The C2 domains of synaptotagmin--partners in exocytosis. , 2004, Trends in biochemical sciences.

[19]  Tomohiko Maehama,et al.  Crystal Structure of the PTEN Tumor Suppressor Implications for Its Phosphoinositide Phosphatase Activity and Membrane Association , 1999, Cell.

[20]  D. Sabatini,et al.  Hydrolysis of GTP on rab11 is required for the direct delivery of transferrin from the pericentriolar recycling compartment to the cell surface but not from sorting endosomes. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  H. Stenmark,et al.  Phosphatidylinositol 3-phosphate is found in microdomains of early endosomes , 2003, Histochemistry and Cell Biology.

[22]  Wonhwa Cho,et al.  Membrane-binding and activation mechanism of PTEN , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[23]  P. Majerus,et al.  SopB, a protein required for virulence of Salmonella dublin, is an inositol phosphate phosphatase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  W. Frankel,et al.  A Null Mutation in Inositol Polyphosphate 4-Phosphatase Type I Causes Selective Neuronal Loss in Weeble Mutant Mice , 2001, Neuron.

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

[26]  S. Emr,et al.  The role of phosphoinositides in membrane transport. , 2001, Current opinion in cell biology.

[27]  P. Hawkins,et al.  Agonist-stimulated synthesis of phosphatidylinositol(3,4,5)-trisphosphate: a new intracellular signalling system? , 1993, Biochimica et biophysica acta.

[28]  J. van der Kaay,et al.  Distinct Phosphatidylinositol 3-Kinase Lipid Products Accumulate upon Oxidative and Osmotic Stress and Lead to Different Cellular Responses* , 1999, The Journal of Biological Chemistry.

[29]  F. Maxfield,et al.  Endocytic recycling , 2004, Nature Reviews Molecular Cell Biology.

[30]  C. Mitchell,et al.  Sec15 Is an Effector for the Rab11 GTPase in Mammalian Cells* , 2004, Journal of Biological Chemistry.

[31]  M. Yaffe,et al.  The PX domains of p47phox and p40phox bind to lipid products of PI(3)K , 2001, Nature Cell Biology.

[32]  M. Lemmon,et al.  Phosphoinositide Recognition Domains , 2003, Traffic.

[33]  Lewis C Cantley,et al.  The phosphoinositide 3-kinase pathway. , 2002, Science.

[34]  C. Mitchell,et al.  The SH2-containing inositol polyphosphate 5-phosphatase, SHIP-2, binds filamin and regulates submembraneous actin , 2001, The Journal of cell biology.

[35]  R. Atkins,et al.  Inositol Polyphosphate 4-Phosphatase Is Inactivated by Calpain-mediated Proteolysis in Stimulated Human Platelets* , 1997, The Journal of Biological Chemistry.

[36]  M. Waterfield,et al.  The Class II Phosphoinositide 3-Kinase PI3K-C2α Is Concentrated in the Trans-Golgi Network and Present in Clathrin-coated Vesicles* , 2000, The Journal of Biological Chemistry.

[37]  M. Lemmon,et al.  Structural basis for discrimination of 3-phosphoinositides by pleckstrin homology domains. , 2000, Molecular cell.

[38]  I. Mills,et al.  Inhibition of endosome fusion by wortmannin persists in the presence of activated Rab5. , 1998, Molecular biology of the cell.

[39]  E. Smythe Direct interactions between rab GTPases and cargo. , 2002, Molecular cell.

[40]  A. Klippel,et al.  Evidence for phosphatidylinositol 3-kinase as a regulator of endocytosis via activation of Rab5. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[41]  X. Chen,et al.  Regulation of intracellular trafficking of the EGF receptor by Rab5 in the absence of phosphatidylinositol 3‐kinase activity , 2001, EMBO reports.

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

[43]  P. Stahl,et al.  Characterization of Rab5:Q79L-stimulated endosome fusion. , 1996, Archives of biochemistry and biophysics.

[44]  G. Prestwich,et al.  Phosphatidylinositol (3,4,5)P3 Is Essential but Not Sufficient for Protein Kinase B (PKB) Activation; Phosphatidylinositol (3,4)P2 Is Required for PKB Phosphorylation at Ser-473 , 2002, The Journal of Biological Chemistry.

[45]  J. Salamero,et al.  Rab11 Regulates the Compartmentalization of Early Endosomes Required for Efficient Transport from Early Endosomes to the Trans-Golgi Network , 2000, The Journal of cell biology.

[46]  J. Backer,et al.  Vps34p differentially regulates endocytosis from the apical and basolateral domains in polarized hepatic cells , 2001, The Journal of cell biology.

[47]  M. Czech Dynamics of phosphoinositides in membrane retrieval and insertion. , 2003, Annual review of physiology.

[48]  J. Whisstock,et al.  Identification of a Novel Domain in Two Mammalian Inositol-polyphosphate 5-Phosphatases That Mediates Membrane Ruffle Localization , 2003, The Journal of Biological Chemistry.

[49]  J. Hurley,et al.  Subcellular targeting by membrane lipids. , 2001, Current opinion in cell biology.

[50]  Ching-shih Chen,et al.  A Novel Integrin-activated Pathway Forms PKB/Akt- stimulatory Phosphatidylinositol 3,4-Bisphosphate via Phosphatidylinositol 3-Phosphate in Platelets* , 1998, The Journal of Biological Chemistry.