Identification and Characterization of an Essential Family of Inositol Polyphosphate 5-Phosphatases (INP51, INP52 and INP53 Gene Products) in the Yeast Saccharomyces cerevisiae

Corresponding author: John D. York, Department of Pharmacology and Cancer Biology, Duke University Medical Center, DUMC 3813, Durham, NC 27710, yorkj{at}acpub.duke.edu (E-mail).

Communicating editor: M. JOHNSTON

We recently demonstrated that the S. cerevisiae INP51 locus (YIL002c) encodes an inositol polyphosphate 5-phosphatase. Here we describe two related yeast loci, INP52 (YNL106c) and INP53 (YOR109w). Like Inp51p, the primary structures of Inp52p and Inp53p resemble the mammalian synaptic vesicle-associated protein, synaptojanin, and contain a carboxy-terminal catalytic domain and an amino-terminal SAC1-like segment. Inp51p (108 kD), Inp52p (136 kD) and Inp53p (124 kD) are membrane-associated. Single null mutants (inp51, inp52, or inp53) are viable. Both inp51 inp52 and inp52 inp53 double mutants display compromised cell growth, whereas an inp51 inp53 double mutant does not. An inp51 inp52 inp53 triple mutant is inviable on standard medium, but can grow weakly on media supplemented with an osmotic stabilizer (1 M sorbitol). An inp51 mutation, and to a lesser degree an inp52 mutation, confers cold-resistant growth in a strain background that cannot grow at temperatures below 15°. Analysis of inositol metabolites in vivo showed measurable accumulation of phosphatidylinositol 4,5-bisphosphate in the inp51 mutant. Electron microscopy revealed plasma membrane invaginations and cell wall thickening in double mutants and the triple mutant grown in sorbitol-containing medium. A fluorescent dye that detects endocytic and vacuolar membranes suggests that the vacuole is highly fragmented in inp51 inp52 double mutants. Our observations indicate that Inp51p, Inp52p, and Inp53p have distinct functions and that substrates and/or products of inositol polyphosphate 5-phosphatases may have roles in vesicle trafficking, membrane structure, and/or cell wall formation.

This article has been cited by other articles:

				H. Badrane, M. H. Nguyen, S. Cheng, V. Kumar, H. Derendorf, K. A. Iczkowski, and C. J. Clancy The Candida albicans phosphatase Inp51p interacts with the EH domain protein Irs4p, regulates phosphatidylinositol-4,5-bisphosphate levels and influences hyphal formation, the cell integrity pathway and virulence Microbiology, November 1, 2008; 154(11): 3296 - 3308. [Abstract] [Full Text] [PDF]

				M. Umemura, M. Fujita, T. Yoko-o, A. Fukamizu, and Y. Jigami Saccharomyces cerevisiae CWH43 Is Involved in the Remodeling of the Lipid Moiety of GPI Anchors to Ceramides Mol. Biol. Cell, November 1, 2007; 18(11): 4304 - 4316. [Abstract] [Full Text] [PDF]

				F. Wiradjaja, L. M. Ooms, S. Tahirovic, E. Kuhne, R. J. Devenish, A. L. Munn, R. C. Piper, P. Mayinger, and C. A. Mitchell Inactivation of the Phosphoinositide Phosphatases Sac1p and Inp54p Leads to Accumulation of Phosphatidylinositol 4,5-Bisphosphate on Vacuole Membranes and Vacuolar Fusion Defects J. Biol. Chem., June 1, 2007; 282(22): 16295 - 16307. [Abstract] [Full Text] [PDF]

				Y. Sun, S. Carroll, M. Kaksonen, J. Y. Toshima, and D. G. Drubin PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization J. Cell Biol., April 23, 2007; 177(2): 355 - 367. [Abstract] [Full Text] [PDF]

				W. R. Parrish, C. J. Stefan, and S. D. Emr PtdIns(3)P accumulation in triple lipid-phosphatase-deletion mutants triggers lethal hyperactivation of the Rho1p/Pkc1p cell-integrity MAP kinase pathway J. Cell Sci., December 1, 2005; 118(23): 5589 - 5601. [Abstract] [Full Text] [PDF]

				Y.-M. Zhang, C. O. Rock, and S. Jackowski Feedback Regulation of Murine Pantothenate Kinase 3 by Coenzyme A and Coenzyme A Thioesters J. Biol. Chem., September 23, 2005; 280(38): 32594 - 32601. [Abstract] [Full Text] [PDF]

				A. M. Seeds, R. J. Bastidas, and J. D. York Molecular Definition of a Novel Inositol Polyphosphate Metabolic Pathway Initiated by Inositol 1,4,5-Trisphosphate 3-Kinase Activity in Saccharomyces cerevisiae J. Biol. Chem., July 29, 2005; 280(30): 27654 - 27661. [Abstract] [Full Text] [PDF]

				C. J. Stefan, S. M. Padilla, A. Audhya, and S. D. Emr The Phosphoinositide Phosphatase Sjl2 Is Recruited to Cortical Actin Patches in the Control of Vesicle Formation and Fission during Endocytosis Mol. Cell. Biol., April 15, 2005; 25(8): 2910 - 2923. [Abstract] [Full Text] [PDF]

				M. Fadri, A. Daquinag, S. Wang, T. Xue, and J. Kunz The Pleckstrin Homology Domain Proteins Slm1 and Slm2 Are Required for Actin Cytoskeleton Organization in Yeast and Bind Phosphatidylinositol-4,5-Bisphosphate and TORC2 Mol. Biol. Cell, April 1, 2005; 16(4): 1883 - 1900. [Abstract] [Full Text] [PDF]

				S. J. York, B. N. Armbruster, P. Greenwell, T. D. Petes, and J. D. York Inositol Diphosphate Signaling Regulates Telomere Length J. Biol. Chem., February 11, 2005; 280(6): 4264 - 4269. [Abstract] [Full Text] [PDF]

				Y. Sun, M. Kaksonen, D. T. Madden, R. Schekman, and D. G. Drubin Interaction of Sla2p's ANTH Domain with PtdIns(4,5)P2 Is Important for Actin-dependent Endocytic Internalization Mol. Biol. Cell, February 1, 2005; 16(2): 717 - 730. [Abstract] [Full Text] [PDF]

				H. Morales-Johansson, P. Jenoe, F. T. Cooke, and M. N. Hall Negative Regulation of Phosphatidylinositol 4,5-Bisphosphate Levels by the INP51-associated Proteins TAX4 and IRS4 J. Biol. Chem., September 17, 2004; 279(38): 39604 - 39610. [Abstract] [Full Text] [PDF]

				W. R. Parrish, C. J. Stefan, and S. D. Emr Essential Role for the Myotubularin-related Phosphatase Ymr1p and the Synaptojanin-like Phosphatases Sjl2p and Sjl3p in Regulation of Phosphatidylinositol 3-Phosphate in Yeast Mol. Biol. Cell, August 1, 2004; 15(8): 3567 - 3579. [Abstract] [Full Text] [PDF]

				M. G. Roth Phosphoinositides in Constitutive Membrane Traffic Physiol Rev, July 1, 2004; 84(3): 699 - 730. [Abstract] [Full Text] [PDF]

				S. A. Rudge, D. M. Anderson, and S. D. Emr Vacuole Size Control: Regulation of PtdIns(3,5)P2 Levels by the Vacuole-associated Vac14-Fig4 Complex, a PtdIns(3,5)P2-specific Phosphatase Mol. Biol. Cell, January 1, 2004; 15(1): 24 - 36. [Abstract] [Full Text] [PDF]

				S.-A. Ha, J. Torabinejad, D. B. DeWald, M. R. Wenk, L. Lucast, P. De Camilli, R. A. Newitt, R. Aebersold, and S. F. Nothwehr The Synaptojanin-like Protein Inp53/Sjl3 Functions with Clathrin in a Yeast TGN-to-Endosome Pathway Distinct from the GGA Protein-dependent Pathway Mol. Biol. Cell, April 1, 2003; 14(4): 1319 - 1333. [Abstract] [Full Text] [PDF]

				H. M. Loovers, K. Veenstra, H. Snippe, X. Pesesse, C. Erneux, and P. J. M. van Haastert A Diverse Family of Inositol 5-Phosphatases Playing a Role in Growth and Development in Dictyostelium discoideum J. Biol. Chem., February 14, 2003; 278(8): 5652 - 5658. [Abstract] [Full Text] [PDF]

				E. Dubois, B. Scherens, F. Vierendeels, M. M. W. Ho, F. Messenguy, and S. B. Shears In Saccharomyces cerevisiae, the Inositol Polyphosphate Kinase Activity of Kcs1p Is Required for Resistance to Salt Stress, Cell Wall Integrity, and Vacuolar Morphogenesis J. Biol. Chem., June 21, 2002; 277(26): 23755 - 23763. [Abstract] [Full Text] [PDF]

				Y. K. Bui and P. W. Sternberg Caenorhabditis elegans Inositol 5-Phosphatase Homolog Negatively Regulates Inositol 1,4,5-Triphosphate Signaling in Ovulation Mol. Biol. Cell, May 1, 2002; 13(5): 1641 - 1651. [Abstract] [Full Text] [PDF]

				G. Konrad, T. Schlecker, F. Faulhammer, and P. Mayinger Retention of the Yeast Sac1p Phosphatase in the Endoplasmic Reticulum Causes Distinct Changes in Cellular Phosphoinositide Levels and Stimulates Microsomal ATP Transport J. Biol. Chem., March 15, 2002; 277(12): 10547 - 10554. [Abstract] [Full Text] [PDF]

				E. S. Seeley, M. Kato, N. Margolis, W. Wickner, and G. Eitzen Genomic Analysis of Homotypic Vacuole Fusion Mol. Biol. Cell, March 1, 2002; 13(3): 782 - 794. [Abstract] [Full Text] [PDF]

				S.-A. Ha, J. T. Bunch, H. Hama, D. B. DeWald, and S. F. Nothwehr A Novel Mechanism for Localizing Membrane Proteins to Yeast Trans-Golgi Network Requires Function of Synaptojanin-like Protein Mol. Biol. Cell, October 1, 2001; 12(10): 3175 - 3190. [Abstract] [Full Text] [PDF]

				M. Foti, A. Audhya, and S. D. Emr Sac1 Lipid Phosphatase and Stt4 Phosphatidylinositol 4-Kinase Regulate a Pool of Phosphatidylinositol 4-Phosphate That Functions in the Control of the Actin Cytoskeleton and Vacuole Morphology Mol. Biol. Cell, August 1, 2001; 12(8): 2396 - 2411. [Abstract] [Full Text] [PDF]

				S. E. Berdy, J. Kudla, W. Gruissem, and G. E. Gillaspy Molecular Characterization of At5PTase1, an Inositol Phosphatase Capable of Terminating Inositol Trisphosphate Signaling Plant Physiology, June 1, 2001; 126(2): 801 - 810. [Abstract] [Full Text] [PDF]

				L. M. Ooms, B. K. McColl, F. Wiradjaja, A. P. W. Wijayaratnam, P. Gleeson, M. J. Gething, J. Sambrook, and C. A. Mitchell The Yeast Inositol Polyphosphate 5-Phosphatases Inp52p and Inp53p Translocate to Actin Patches following Hyperosmotic Stress: Mechanism for Regulating Phosphatidylinositol 4,5-Bisphosphate at Plasma Membrane Invaginations Mol. Cell. Biol., December 15, 2000; 20(24): 9376 - 9390. [Abstract] [Full Text] [PDF]

				T. W. Harris, E. Hartwieg, H. R. Horvitz, and E. M. Jorgensen Mutations in Synaptojanin Disrupt Synaptic Vesicle Recycling J. Cell Biol., August 7, 2000; 150(3): 589 - 600. [Abstract] [Full Text] [PDF]

				A. Mayer, D. Scheglmann, S. Dove, A. Glatz, W. Wickner, and A. Haas Phosphatidylinositol 4,5-Bisphosphate Regulates Two Steps of Homotypic Vacuole Fusion Mol. Biol. Cell, March 1, 2000; 11(3): 807 - 817. [Abstract] [Full Text]

				E. S. Bensen, G. Costaguta, and G. S. Payne Synthetic Genetic Interactions With Temperature-Sensitive Clathrin in Saccharomyces cerevisiae: Roles for Synaptojanin-Like Inp53p and Dynamin-Related Vps1p in Clathrin-Dependent Protein Sorting at the trans-Golgi Network Genetics, January 1, 2000; 154(1): 83 - 97. [Abstract] [Full Text]

				H. Hama, E. A. Schnieders, J. Thorner, J. Y. Takemoto, and D. B. DeWald Direct Involvement of Phosphatidylinositol 4-Phosphate in Secretion in the Yeast Saccharomyces cerevisiae J. Biol. Chem., November 26, 1999; 274(48): 34294 - 34300. [Abstract] [Full Text] [PDF]

				D. A. Thompson and F. W. Stahl Genetic Control of Recombination Partner Preference in Yeast Meiosis: Isolation and Characterization of Mutants Elevated for Meiotic Unequal Sister-Chromatid Recombination Genetics, October 1, 1999; 153(2): 621 - 641. [Abstract] [Full Text] [PDF]

				S. Guo, L. E. Stolz, S. M. Lemrow, and J. D. York SAC1-like Domains of Yeast SAC1, INP52, and INP53 and of Human Synaptojanin Encode Polyphosphoinositide Phosphatases J. Biol. Chem., May 7, 1999; 274(19): 12990 - 12995. [Abstract] [Full Text] [PDF]

				B. D. Spiegelberg, J.-P. Xiong, J. J. Smith, R. F. Gu, and J. D. York Cloning and Characterization of a Mammalian Lithium-sensitive Bisphosphate 3'-Nucleotidase Inhibited by Inositol 1,4-Bisphosphate J. Biol. Chem., May 7, 1999; 274(19): 13619 - 13628. [Abstract] [Full Text] [PDF]

				P. W. Majerus, M. V. Kisseleva, and F. A. Norris The Role of Phosphatases in Inositol Signaling Reactions J. Biol. Chem., April 16, 1999; 274(16): 10669 - 10672. [Full Text] [PDF]

				J. D. Gary, A. E. Wurmser, C. J. Bonangelino, L. S. Weisman, and S. D. Emr Fab1p Is Essential for PtdIns(3)P 5-Kinase Activity and the Maintenance of Vacuolar Size and Membrane Homeostasis J. Cell Biol., October 5, 1998; 143(1): 65 - 79. [Abstract] [Full Text] [PDF]

				B Singer-Kruger, Y Nemoto, L Daniell, S Ferro-Novick, and P De Camilli Synaptojanin family members are implicated in endocytic membrane traffic in yeast J. Cell Sci., January 11, 1998; 111(22): 3347 - 3356. [Abstract] [PDF]

				F. Wiradjaja, L. M. Ooms, J. C. Whisstock, B. McColl, L. Helfenbaum, J. F. Sambrook, M.-J. Gething, and C. A. Mitchell The Yeast Inositol Polyphosphate 5-Phosphatase Inp54p Localizes to the Endoplasmic Reticulum via a C-terminal Hydrophobic Anchoring Tail. REGULATION OF SECRETION FROM THE ENDOPLASMIC RETICULUM J. Biol. Chem., March 2, 2001; 276(10): 7643 - 7653. [Abstract] [Full Text] [PDF]

				Y. Nemoto, M. R. Wenk, M. Watanabe, L. Daniell, T. Murakami, N. Ringstad, H. Yamada, K. Takei, and P. De Camilli Identification and Characterization of a Synaptojanin 2 Splice Isoform Predominantly Expressed in Nerve Terminals J. Biol. Chem., October 26, 2001; 276(44): 41133 - 41142. [Abstract] [Full Text] [PDF]

				C. J. Stefan, A. Audhya, and S. D. Emr The Yeast Synaptojanin-like Proteins Control the Cellular Distribution of Phosphatidylinositol (4,5)-Bisphosphate Mol. Biol. Cell, February 1, 2002; 13(2): 542 - 557. [Abstract] [Full Text] [PDF]

				J. D. Gary, T. K. Sato, C. J. Stefan, C. J. Bonangelino, L. S. Weisman, and S. D. Emr Regulation of Fab1 Phosphatidylinositol 3-Phosphate 5-Kinase Pathway by Vac7 Protein and Fig4, a Polyphosphoinositide Phosphatase Family Member Mol. Biol. Cell, April 1, 2002; 13(4): 1238 - 1251. [Abstract] [Full Text] [PDF]