Regulation of Phagocytosis in Dictyostelium by the Inositol 5‐Phosphatase OCRL Homolog Dd5P4

Phosphoinositides are involved in endocytosis in both mammalian cells and the amoeba Dictyostelium discoideum. Dd5P4 is the Dictyostelium homolog of human OCRL (oculocerebrorenal syndrome of Lowe); both have a RhoGAP domain and a 5‐phosphatase domain that acts on phosphatidylinositol 4,5‐bisphosphate/phosphatidylinositol 3,4,5‐trisphosphate (PI(3,4,5)P3). Inactivation of Dd5P4 inhibits growth on liquid medium and on bacteria. Dd5p4‐null cells are impaired in phagocytosis of yeast cells. In wild‐type cells, PI(3,4,5)P3 is formed and converted to PI(3,4)P2 just before closure of the phagocytic cup. In dd5p4‐null cells, a phagocytic cup is formed upon contact with the yeast cell, and PI(3,4,5)P3 is still produced, but the phagocytic cup does not close. We suggest that Dd5P4 regulates the conversion of PI(3,4,5)P3 to PI(3,4)P2 and that this conversion is essential for closure of the phagocytic cup. Phylogenetic analysis of OCRL‐like 5‐phosphatases with RhoGAP domains reveal that D. discoideum Dd5P4 is a surprisingly close homolog of human OCRL, the protein responsible for Lowe syndrome. We expressed human OCRL in dd5p4‐null cells. Growth on bacteria and axenic medium is largely restored, whereas the rate of phagocytosis of yeast cells is partly restored, indicating that human OCRL can functionally replace Dictyostelium Dd5P4.

[1]  Takehiko Sasaki,et al.  [Phosphoinositides in membrane traffic]. , 2008, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[2]  E. Eisenberg,et al.  Lowe syndrome protein OCRL1 interacts with clathrin and regulates protein trafficking between endosomes and the trans-Golgi network. , 2005, Molecular biology of the cell.

[3]  J. Lunardi,et al.  Lowe syndrome protein Ocrl1 is translocated to membrane ruffles upon Rac GTPase activation: a new perspective on Lowe syndrome pathophysiology. , 2005, Human molecular genetics.

[4]  D. Dormann,et al.  In vivo analysis of 3-phosphoinositide dynamics during Dictyostelium phagocytosis and chemotaxis , 2004, Journal of Cell Science.

[5]  P. Majerus,et al.  The inositol polyphosphate 5-phosphatase Ocrl associates with endosomes that are partially coated with clathrin. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Dixon,et al.  A PTEN-related 5-Phosphatidylinositol Phosphatase Localized in the Golgi* , 2003, Journal of Biological Chemistry.

[7]  J. Lunardi,et al.  Lowe syndrome protein OCRL1 interacts with Rac GTPase in the trans-Golgi network. , 2003, Human molecular genetics.

[8]  R. Nussbaum,et al.  Phosphoinositide profiling in complex lipid mixtures using electrospray ionization mass spectrometry , 2003, Nature Biotechnology.

[9]  P. Devreotes,et al.  Eukaryotic Chemotaxis: Distinctions between Directional Sensing and Polarization* , 2003, Journal of Biological Chemistry.

[10]  P. V. van Haastert,et al.  A Diverse Family of Inositol 5-Phosphatases Playing a Role in Growth and Development in Dictyostelium discoideum * 210 , 2003, The Journal of Biological Chemistry.

[11]  R. Nussbaum,et al.  The deficiency of PIP2 5-phosphatase in Lowe syndrome affects actin polymerization. , 2002, American journal of human genetics.

[12]  Richard A. Firtel,et al.  Spatial and Temporal Regulation of 3-Phosphoinositides by PI 3-Kinase and PTEN Mediates Chemotaxis , 2002, Cell.

[13]  P. Devreotes,et al.  Tumor Suppressor PTEN Mediates Sensing of Chemoattractant Gradients , 2002, Cell.

[14]  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.

[15]  Roger L. Williams.  Faculty Opinions recommendation of Restricted accumulation of phosphatidylinositol 3-kinase products in a plasmalemmal subdomain during Fc gamma receptor-mediated phagocytosis. , 2001 .

[16]  S. Grinstein,et al.  Restricted Accumulation of Phosphatidylinositol 3-Kinase Products in a Plasmalemmal Subdomain during Fcγ Receptor-Mediated Phagocytosis , 2001, The Journal of cell biology.

[17]  James H. Hurley,et al.  Specificity Determinants in Phosphoinositide Dephosphorylation Crystal Structure of an Archetypal Inositol Polyphosphate 5-Phosphatase , 2001, Cell.

[18]  J. Cardelli,et al.  Phagocytosis and Macropinocytosis in Dictyostelium: Phosphoinositide‐Based Processes, Biochemically Distinct , 2001, Traffic.

[19]  J. Sambrook,et al.  Mammalian inositol polyphosphate 5-phosphatase II can compensate for the absence of all three yeast Sac1-like-domain-containing 5-phosphatases. , 2001, The Biochemical journal.

[20]  C. Helgason,et al.  A Regulatory Role for Src Homology 2 Domain–Containing Inositol 5′-Phosphatase (Ship) in Phagocytosis Mediated by Fcγ Receptors and Complement Receptor 3 (αMβ2; Cd11b/Cd18) , 2001, The Journal of experimental medicine.

[21]  S. Grinstein,et al.  Localized Biphasic Changes in Phosphatidylinositol-4,5-Bisphosphate at Sites of Phagocytosis , 2000, The Journal of cell biology.

[22]  J. Faix,et al.  Rac1 GTPases control filopodia formation, cell motility, endocytosis, cytokinesis and development in Dictyostelium. , 2000, Journal of cell science.

[23]  P. V. Haastert,et al.  Optimizing heterologous expression in dictyostelium: importance of 5' codon adaptation. , 2000, Nucleic acids research.

[24]  R. Nussbaum,et al.  Ocrl1, a PtdIns(4,5)P2 5-Phosphatase, Is Localized to the Trans-Golgi Network of Fibroblasts and Epithelial Cells , 2000, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[25]  A. Ungewickell,et al.  Increased levels of plasma lysosomal enzymes in patients with Lowe syndrome. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. McCormick,et al.  Essential Role of Phosphoinositide Metabolism in Synaptic Vesicle Recycling , 1999, Cell.

[27]  X. Pesesse,et al.  The diversity and possible functions of the inositol polyphosphate 5-phosphatases. , 1998, Biochimica et biophysica acta.

[28]  S. Bozzaro,et al.  G Protein β Subunit–null Mutants Are Impaired in Phagocytosis and Chemotaxis Due to Inappropriate Regulation of the Actin Cytoskeleton , 1998, The Journal of cell biology.

[29]  E. Philip,et al.  Cell Lines from Kidney Proximal Tubules of a Patient with Lowe Syndrome Lack OCRL Inositol Polyphosphate 5-Phosphatase and Accumulate Phosphatidylinositol 4,5-Bisphosphate* , 1998, The Journal of Biological Chemistry.

[30]  K. Zhou,et al.  Disruption of Dictyostelium PI3K genes reduces [32P]phosphatidylinositol 3,4 bisphosphate and [32P]phosphatidylinositol trisphosphate levels, alters F-actin distribution and impairs pinocytosis. , 1998, Journal of cell science.

[31]  Katrin Rittinger,et al.  Structure at 1.65 Å of RhoA and its GTPase-activating protein in complex with a transition-state analogue , 1997, Nature.

[32]  J. Bush,et al.  Inactivation of Two Dictyostelium discoideum Genes, DdPIK1 and DdPIK2, Encoding Proteins Related to Mammalian Phosphatidylinositide 3-kinases, Results in Defects in Endocytosis, Lysosome to Postlysosome Transport, and Actin Cytoskeleton Organization , 1997, The Journal of cell biology.

[33]  P. Majerus,et al.  Mutation of the conserved domains of two inositol polyphosphate 5-phosphatases. , 1996, Biochemistry.

[34]  G. Gerisch,et al.  Coronin involved in phagocytosis: Dynamics of particle-induced relocalization visualized by a green fluorescent protein tag , 1995, Cell.

[35]  S. Suchy,et al.  Lowe syndrome, a deficiency of phosphatidylinositol 4,5-bisphosphate 5-phosphatase in the Golgi apparatus. , 1995, Human molecular genetics.

[36]  G. Mayr,et al.  Role of phospholipase C in Dictyostelium: formation of inositol 1,4,5‐trisphosphate and normal development in cells lacking phospholipase C activity. , 1994, The EMBO journal.