Colocalisation of the protein tyrosine phosphatases PTP-SL and PTPBR7 with β4-adaptin in neuronal cells

The mouse gene Ptprr encodes the neuronal protein tyrosine phosphatases PTP-SL and PTPBR7. These proteins differ in their N-terminal domains, with PTP-SL being a cytosolic, membrane-associated phosphatase and PTPBR7 a type I transmembrane protein. In this study, we further explored the nature of the PTP-SL-associated vesicles in neuronal cells using a panel of organelle markers and noted a comparable subcellular distribution for PTP-SL and the β4-adaptin subunit of the AP4 complex. PTP-SL, PTPBR7 and β4-adaptin are localised at the Golgi apparatus and at vesicles throughout the cytoplasm. Immunohistochemical analysis demonstrated that PTP-SL, PTPBR7 and β4-adaptin are all endogenously expressed in brain. Interestingly, coexpression of PTP-SL and β4-adaptin leads to an altered subcellular localisation for PTP-SL. Instead of the Golgi and vesicle-type staining pattern, still observable for β4-adaptin, PTP-SL is now distributed throughout the cytoplasm. Although β4-adaptin was found to interact with the phosphatase domain of PTP-SL and PTPBR7 in the yeast two-hybrid system, it failed to do so in transfected neuronal cells. Our data suggest that the tyrosine phosphatases PTP-SL and PTPBR7 may be involved in the formation and transport of AP4-coated vesicles or in the dephosphorylation of their transmembrane cargo molecules at or near the Golgi apparatus.

[1]  D. Rotin,et al.  The Second Catalytic Domain of Protein Tyrosine Phosphatase δ (PTPδ) Binds to and Inhibits the First Catalytic Domain of PTPς , 1998, Molecular and Cellular Biology.

[2]  M. Isobe,et al.  Molecular cloning and chromosomal mapping of a human protein-tyrosine phosphatase LC-PTP. , 1992, Biochemical and biophysical research communications.

[3]  L. Butcher,et al.  Deficient LAR expression decreases basal forebrain cholinergic neuronal size and hippocampal cholinergic innervation , 1997, Journal of neuroscience research.

[4]  B. Goud,et al.  The small GTPase Rab6B, a novel Rab6 subfamily member, is cell-type specifically expressed and localised to the Golgi apparatus. , 2000, Journal of cell science.

[5]  Y. Watanabe,et al.  Cloning and expression of PCPTP1 encoding protein tyrosine phosphatase. , 1995, Gene.

[6]  R. Parton,et al.  A lipid associated with the antiphospholipid syndrome regulates endosome structure and function , 1998, Nature.

[7]  J. Fransen,et al.  The mouse Ptprr gene encodes two protein tyrosine phosphatases, PTP‐SL and PTPBR7, that display distinct patterns of expression during neural development , 1999, The European journal of neuroscience.

[8]  M. Minden,et al.  Cloning and expression of an inducible lymphoid‐specific, protein tyrosine phosphatase (HePTPase) , 1992, European journal of immunology.

[9]  M. Robinson,et al.  Coats and vesicle budding. , 1997, Trends in cell biology.

[10]  H. Hauri,et al.  Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus , 1988, The Journal of cell biology.

[11]  B. Kari,et al.  Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes , 1986, Journal of virology.

[12]  K. Mikoshiba,et al.  Impaired learning with enhanced hippocampal long‐term potentiation in PTPδ‐deficient mice , 2000 .

[13]  A. Wilde,et al.  In vivo phosphorylation of adaptors regulates their interaction with clathrin , 1996, The Journal of cell biology.

[14]  H. Hameister,et al.  Molecular cloning of a mouse epithelial protein‐tyrosine phosphatase with similarities to submembranous proteins , 1995, Journal of cellular biochemistry.

[15]  Dynein, Dynactin, and Kinesin II's Interaction with Microtubules Is Regulated during Bidirectional Organelle Transport , 2000, The Journal of cell biology.

[16]  M. Klagsbrun,et al.  Heparin-like Molecules on the Cell Surface Potentiate Binding of Diphtheria Toxin to the Diphtheria Toxin Receptor/Membrane-anchored Heparin-binding Epidermal Growth Factor-like Growth Factor (*) , 1995, The Journal of Biological Chemistry.

[17]  G. Church,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[18]  C. Goodman,et al.  The Transmembrane Tyrosine Phosphatase DLAR Controls Motor Axon Guidance in Drosophila , 1996, Cell.

[19]  P. Wahle,et al.  Cellular and molecular characterization of a brain-enriched protein tyrosine phosphatase , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[20]  J. Naegele,et al.  STEP61: A Member of a Family of Brain-Enriched PTPs Is Localized to the Endoplasmic Reticulum , 1996, The Journal of Neuroscience.

[21]  Tomas Mustelin,et al.  Crosstalk between cAMP-dependent kinase and MAP kinase through a protein tyrosine phosphatase , 1999, Nature Cell Biology.

[22]  J. Bonifacino,et al.  Cloning, expression, and localization of a novel γ‐adaptin‐like molecule , 1998 .

[23]  Carmen Blanco-Aparicio,et al.  A Novel Regulatory Mechanism of Map Kinases Activation and Nuclear Translocation Mediated by Pka and the Ptp-Sl Tyrosine Phosphatase , 1999, The Journal of cell biology.

[24]  H. Hauri,et al.  Immuno-electronmicroscopical localization of a microvillus membrane disaccharidase in the human small-intestinal epithelium with monoclonal antibodies. , 1985, European journal of cell biology.

[25]  M. Zerial,et al.  Membrane Dynamics in Endocytosis , 1996, Cell.

[26]  Peter J Verveer,et al.  Imaging Sites of Receptor Dephosphorylation by PTP1B on the Surface of the Endoplasmic Reticulum , 2002, Science.

[27]  Roger Brent,et al.  C dil, a Human Gl and S Phase Protein Phosphatase That Associates with Cdk2 , 2003 .

[28]  T. Hamaoka,et al.  Inactivation of mitogen-activated protein kinases by a mammalian tyrosine-specific phosphatase, PTPBR7. , 1999, Biochemical and biophysical research communications.

[29]  S. Freedman,et al.  Endocytosis at the apical plasma membrane of pancreatic acinar cells is regulated by tyrosine kinases. , 1999, The American journal of physiology.

[30]  B. Wieringa,et al.  A novel receptor-type protein tyrosine phosphatase with a single catalytic domain is specifically expressed in mouse brain. , 1995, The Biochemical journal.

[31]  L. Traub.  Clathrin-associated adaptor proteins - putting it all together. , 1997, Trends in cell biology.

[32]  P. Nucifora,et al.  Tyrosine Phosphorylation Regulates Rapid Endocytosis in Adrenal Chromaffin Cells , 1999, The Journal of Neuroscience.

[33]  P. Lombroso,et al.  A Neuronal Protein Tyrosine Phosphatase Induced by Nerve Growth Factor (*) , 1995, The Journal of Biological Chemistry.

[34]  W. Hendriks,et al.  A decrease in size and number of basal forebrain cholinergic neurons is paralleled by diminished hippocampal cholinergic innervation in mice lacking leukocyte common antigen-related protein tyrosine phosphatase activity , 2001, Neuroscience.

[35]  M. Robinson,et al.  Characterization of a fourth adaptor-related protein complex. , 1999, Molecular biology of the cell.

[36]  N. Bucay,et al.  Protein tyrosine phosphatase (PC12, Br7,Sl) family: Expression characterization in the adult human and mouse , 2000 .

[37]  F. Brodsky.  New fashions in vesicle coats. , 1997, Trends in cell biology.

[38]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[39]  D. Rotin,et al.  The second catalytic domain of protein tyrosine phosphatase delta (PTP delta) binds to and inhibits the first catalytic domain of PTP sigma. , 1998, Molecular and cellular biology.

[40]  J. Bonifacino,et al.  AP-4, a Novel Protein Complex Related to Clathrin Adaptors* , 1999, The Journal of Biological Chemistry.

[41]  P. De Camilli,et al.  Role of phosphorylation in regulation of the assembly of endocytic coat complexes. , 1998, Science.

[42]  H. Brinkmann,et al.  Phylogenetic Analysis of Components of the Eukaryotic Vesicle Transport System Reveals a Common Origin of Adaptor Protein Complexes 1, 2, and 3 and the F Subcomplex of the Coatomer COPI , 1999, Journal of Molecular Evolution.

[43]  E. Cuppen,et al.  A FERM domain governs apical confinement of PTP-BL in epithelial cells. , 1999, Journal of cell science.

[44]  R. Pepperkok,et al.  Partitioning of the Golgi Apparatus during Mitosis in Living HeLa Cells , 1997, The Journal of cell biology.

[45]  R. Pulido,et al.  Interaction of Mitogen-activated Protein Kinases with the Kinase Interaction Motif of the Tyrosine Phosphatase PTP-SL Provides Substrate Specificity and Retains ERK2 in the Cytoplasm* , 1999, The Journal of Biological Chemistry.

[46]  P. Camilli,et al.  Direct interaction of the 170 kDa isoform of synaptojanin 1 with clathrin and with the clathrin adaptor AP-2 , 2000, Current Biology.

[47]  N. Bucay,et al.  Protein tyrosine phosphatase (PC12, Br7,S1) family: expression characterization in the adult human and mouse. , 2000, The Anatomical record.

[48]  E. Cuppen,et al.  PDZ motifs in PTP-BL and RIL bind to internal protein segments in the LIM domain protein RIL. , 1998, Molecular biology of the cell.

[49]  J. Bonifacino,et al.  Adaptins: the final recount. , 2001, Molecular biology of the cell.

[50]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[51]  A. Ullrich,et al.  PTP‐SL and STEP protein tyrosine phosphatases regulate the activation of the extracellular signal‐regulated kinases ERK1 and ERK2 by association through a kinase interaction motif , 1998, The EMBO journal.

[52]  B. Neel,et al.  Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. , 1993, Analytical biochemistry.

[53]  Y. Watanabe,et al.  Cloning of PCPTP1-Ce encoding protein tyrosine phosphatase from the rat cerebellum and its restricted expression in Purkinje cells. , 1998, Brain research. Molecular brain research.

[54]  T. Hamaoka,et al.  cDNA Cloning and Characterization of a Novel Receptor-type Protein Tyrosine Phosphatase Expressed Predominantly in the Brain (*) , 1995, The Journal of Biological Chemistry.

[55]  K. Dong,et al.  Inhibition of Protein-tyrosine Phosphatase Stimulates the Dynamin-dependent Endocytosis of ROMK1* , 2002, The Journal of Biological Chemistry.

[56]  M. Robinson,et al.  Targeting signals and subunit interactions in coated vesicle adaptor complexes , 1995, The Journal of cell biology.

[57]  C. Geisler,et al.  beta2-adaptin is constitutively de-phosphorylated by serine/threonine protein phosphatase PP2A and phosphorylated by a staurosporine-sensitive kinase. , 2000, Biochimica et biophysica acta.

[58]  T. Kirchhausen,et al.  A role for the hinge/ear domain of the β chains in the incorporation of AP complexes into clathrin-coated pits and coated vesicles , 1997, Cellular and Molecular Life Sciences CMLS.

[59]  F. Brodsky,et al.  EGF Receptor Signaling Stimulates SRC Kinase Phosphorylation of Clathrin, Influencing Clathrin Redistribution and EGF Uptake , 1999, Cell.

[60]  Timothy E. Kennedy,et al.  Neuroendocrine dysplasia in mice lacking protein tyrosine phosphatase σ , 1999, Nature Genetics.

[61]  S. Freedman,et al.  Endocytosis at the apical plasma membrane of pancreatic acinar cells is regulated by tyrosine kinases. , 1999, American journal of physiology. Cell physiology.

[62]  K. Zinn,et al.  Tyrosine phosphorylation and axon guidance: of mice and flies , 1997, Current Opinion in Neurobiology.

[63]  A. Stoker,et al.  Protein tyrosine phosphatases and neural development , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.