The subcellular localization of the Niemann-Pick Type C proteins depends on the adaptor complex AP-3

Niemann-Pick Type C (NP-C) disease, caused by mutations in either human NPC1 (hNPC1) or human NPC2 (hNPC2), is characterized by the accumulation of unesterified cholesterol in late endosomes. Although it is known that the NP-C proteins are targeted to late endosomal/lysosomal compartments, their delivery mechanisms have not been fully elucidated. To identify mechanisms regulating NP-C protein localization, we used Saccharomyces cerevisiae, which expresses functional homologs of both NP-C proteins – scNcr1p and scNpc2p. Targeting of scNcr1p to the vacuole was perturbed in AP-3-deficient yeast cells, whereas the delivery of scNpc2p was affected by deficiencies in either AP-3 or GGA. We focused on the role of the AP-3 pathway in the targeting of the mammalian NP-C proteins. We found that, although mouse NPC1 (mNPC1) and hNPC2 co-localize with AP-3 to a similar extent in fibroblasts, hNPC2 preferentially co-localizes with AP-1. Importantly, the targeting of both mammalian NPC1 and NPC2 is dependent on AP-3. Moreover, and consistent with the NP-C proteins playing a role in cholesterol metabolism, AP-3-deficient cells have reduced levels of cholesterol. These results provide information about how the NP-C proteins are targeted to their sites of action and illustrate the possibility that defective sorting of the NP-C proteins along the endocytic route can alter cellular cholesterol.

[1]  B. Roux,et al.  Modeling the structure of the StART domains of MLN64 and StAR proteins in complex with cholesterol1s⃞ Published, JLR Papers in Press, September 21, 2006. , 2006, Journal of Lipid Research.

[2]  Mingjie Zhang,et al.  Identification of the yeast R-SNARE Nyv1p as a novel longin domain-containing protein. , 2006, Molecular biology of the cell.

[3]  B. Wainer,et al.  BLOC-1 complex deficiency alters the targeting of adaptor protein complex-3 cargoes. , 2006, Molecular biology of the cell.

[4]  J. Falcón-Pérez,et al.  BLOC-1 interacts with BLOC-2 and the AP-3 complex to facilitate protein trafficking on endosomes. , 2006, Molecular biology of the cell.

[5]  Adam C. Berger,et al.  Saccharomyces cerevisiae Npc2p Is a Functionally Conserved Homologue of the Human Niemann-Pick Disease Type C 2 Protein, hNPC2 , 2005, Eukaryotic Cell.

[6]  A. Corbett,et al.  A Yeast Model System for Functional Analysis of the Niemann–Pick Type C Protein 1 Homolog, Ncr1p , 2005, Traffic.

[7]  J. Bonifacino,et al.  Role of the endocytic machinery in the sorting of lysosome-associated membrane proteins. , 2005, Molecular biology of the cell.

[8]  J. Vance,et al.  Adenovirus expressing an NPC1‐GFP fusion gene corrects neuronal and nonneuronal defects associated with Niemann pick type C disease , 2005, Journal of neuroscience research.

[9]  P. Camilli,et al.  Phosphatidylinositol-4-Kinase Type II α Is a Component of Adaptor Protein-3-derived Vesicles , 2005 .

[10]  J. Shim,et al.  The AP-3 clathrin-associated complex is essential for embryonic and larval development in Caenorhabditis elegans. , 2005, Molecules and cells.

[11]  A. Munn,et al.  Ncr1p, the Yeast Ortholog of Mammalian Niemann Pick C1 Protein, is Dispensable for Endocytic Transport , 2004, Traffic.

[12]  V. Faundez,et al.  The endo-lysosomal sorting machinery interacts with the intermediate filament cytoskeleton. , 2004, Molecular biology of the cell.

[13]  S. E. Miller,et al.  EpsinR is an adaptor for the SNARE protein Vti1b. , 2004, Molecular biology of the cell.

[14]  Y. Ioannou,et al.  Targeting of NPC1 to Late Endosomes Involves Multiple Signals, Including One Residing within the Putative Sterol-sensing Domain* , 2004, Journal of Biological Chemistry.

[15]  R. Lüllmann-Rauch,et al.  Disturbed cholesterol traffic but normal proteolytic function in LAMP-1/LAMP-2 double-deficient fibroblasts. , 2004, Molecular biology of the cell.

[16]  M. Gearing,et al.  AP-3-dependent Mechanisms Control the Targeting of a Chloride Channel (ClC-3) in Neuronal and Non-neuronal Cells* , 2004, Journal of Biological Chemistry.

[17]  D. James,et al.  Combinatorial SNARE complexes with VAMP7 or VAMP8 define different late endocytic fusion events , 2004, EMBO reports.

[18]  M. Robinson.  Adaptable adaptors for coated vesicles. , 2004, Trends in cell biology.

[19]  R. Scheller,et al.  Localization of the AP-3 adaptor complex defines a novel endosomal exit site for lysosomal membrane proteins , 2004, The Journal of cell biology.

[20]  Y. Liu,et al.  Mutagenesis of the putative sterol-sensing domain of yeast Niemann Pick C–related protein reveals a primordial role in subcellular sphingolipid distribution , 2004, The Journal of cell biology.

[21]  A. Levey,et al.  The zinc transporter ZnT3 interacts with AP-3 and it is preferentially targeted to a distinct synaptic vesicle subpopulation. , 2003, Molecular biology of the cell.

[22]  R. Sundaram,et al.  Differential trafficking of the Niemann‐Pick C1 and 2 proteins highlights distinct roles in late endocytic lipid trafficking , 2003, Acta paediatrica (Oslo, Norway : 1992). Supplement.

[23]  J. Bonifacino,et al.  Signals for sorting of transmembrane proteins to endosomes and lysosomes. , 2003, Annual review of biochemistry.

[24]  A. Fayyazi,et al.  Deletion of the SNARE vti1b in Mice Results in the Loss of a Single SNARE Partner, Syntaxin 8 , 2003, Molecular and Cellular Biology.

[25]  M. D'Esposito,et al.  A dual mechanism controlling the localization and function of exocytic v-SNAREs , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[26]  A. Syvänen,et al.  Defective endocytic trafficking of NPC1 and NPC2 underlying infantile Niemann-Pick type C disease. , 2003, Human molecular genetics.

[27]  J. Bonifacino,et al.  Genomic screen for vacuolar protein sorting genes in Saccharomyces cerevisiae. , 2002, Molecular biology of the cell.

[28]  W. Antonin,et al.  The SNAREs vti1a and vti1b have distinct localization and SNARE complex partners. , 2002, European journal of cell biology.

[29]  G. Núñez,et al.  ML -- a conserved domain involved in innate immunity and lipid metabolism. , 2002, Trends in biochemical sciences.

[30]  Alfonso González,et al.  Novel mechanism for regulation of epidermal growth factor receptor endocytosis revealed by protein kinase A inhibition. , 2002, Molecular biology of the cell.

[31]  J. Bonifacino,et al.  Genetic analyses of adaptin function from yeast to mammals. , 2002, Gene.

[32]  R. Schekman,et al.  The yeast clathrin adaptor protein complex 1 is required for the efficient retention of a subset of late Golgi membrane proteins. , 2002, Developmental cell.

[33]  Rachel Rudge,et al.  Assembly and function of AP-3 complexes in cells expressing mutant subunits , 2002, The Journal of cell biology.

[34]  R. Scheller,et al.  The Di-leucine Motif of Vesicle-associated Membrane Protein 4 Is Required for Its Localization and AP-1 Binding* , 2001, The Journal of Biological Chemistry.

[35]  H. Riezman,et al.  End13p/Vps4p is required for efficient transport from early to late endosomes in Saccharomyces cerevisiae. , 2001, Journal of cell science.

[36]  A. Dierich,et al.  The Steroidogenic Acute Regulatory Protein Homolog MLN64, a Late Endosomal Cholesterol-binding Protein* , 2001, The Journal of Biological Chemistry.

[37]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[38]  O. Zhdankina,et al.  Yeast GGA proteins interact with GTP‐bound Arf and facilitate transport through the Golgi , 2001, Yeast.

[39]  R. Wattiaux,et al.  Identification of HE1 as the second gene of Niemann-Pick C disease. , 2000, Science.

[40]  Y. Ioannou,et al.  Transmembrane molecular pump activity of Niemann-Pick C1 protein. , 2000, Science.

[41]  S. Mansour,et al.  Defective organellar membrane protein trafficking in Ap3b1-deficient cells. , 2000, Journal of cell science.

[42]  Yiannis A. Ioannou,et al.  Topological Analysis of Niemann-Pick C1 Protein Reveals That the Membrane Orientation of the Putative Sterol-sensing Domain Is Identical to Those of 3-Hydroxy-3-methylglutaryl-CoA Reductase and Sterol Regulatory Element Binding Protein Cleavage-activating Protein* , 2000, The Journal of Biological Chemistry.

[43]  R. Kelly,et al.  The AP-3 complex required for endosomal synaptic vesicle biogenesis is associated with a casein kinase Ialpha-like isoform. , 2000, Molecular biology of the cell.

[44]  R. Erickson,et al.  Localization of the murine Niemann-Pick C1 protein to two distinct intracellular compartments. , 2000, Journal of lipid research.

[45]  M. Robinson,et al.  A Family of Proteins with γ-Adaptin and Vhs Domains That Facilitate Trafficking between the Trans-Golgi Network and the Vacuole/Lysosome , 2000, The Journal of cell biology.

[46]  L. Liscum.  Niemann–Pick Type C Mutations Cause Lipid Traffic Jam , 2000, Traffic.

[47]  Chunjiang Yu,et al.  Role of Niemann-Pick Type C1 Protein in Intracellular Trafficking of Low Density Lipoprotein-derived Cholesterol* , 2000, The Journal of Biological Chemistry.

[48]  K. von Figura,et al.  The Leucine-based Sorting Motifs in the Cytoplasmic Domain of the Invariant Chain Are Recognized by the Clathrin Adaptors AP1 and AP2 and their Medium Chains* , 1999, The Journal of Biological Chemistry.

[49]  D. Hirsh,et al.  Receptor-mediated endocytosis in the Caenorhabditis elegans oocyte. , 1999, Molecular biology of the cell.

[50]  R. Scheller,et al.  Differential roles of syntaxin 7 and syntaxin 8 in endosomal trafficking. , 1999, Molecular biology of the cell.

[51]  G. Payne,et al.  Adaptor complex-independent clathrin function in yeast. , 1999, Molecular biology of the cell.

[52]  Y. Ioannou,et al.  Niemann-Pick C1 is a late endosome-resident protein that transiently associates with lysosomes and the trans-Golgi network. , 1999, Molecular genetics and metabolism.

[53]  M. Patterson,et al.  The Niemann-Pick C1 Protein Resides in a Vesicular Compartment Linked to Retrograde Transport of Multiple Lysosomal Cargo* , 1999, The Journal of Biological Chemistry.

[54]  T. Ueda,et al.  In vitro binding study of adaptor protein complex (AP-1) to lysosomal targeting motif (LI-motif). , 1999, Biochemical and biophysical research communications.

[55]  H. Watari,et al.  Niemann-Pick C1 protein: obligatory roles for N-terminal domains and lysosomal targeting in cholesterol mobilization. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[56]  D. M. Amundson,et al.  Fluorometric method for the enzymatic determination of cholesterol. , 1999, Journal of biochemical and biophysical methods.

[57]  W. Greene,et al.  A dileucine motif in HIV-1 Nef acts as an internalization signal for CD4 downregulation and binds the AP-1 clathrin adaptor , 1998, Current Biology.

[58]  J. Bonifacino,et al.  Association of the AP-3 adaptor complex with clathrin. , 1998, Science.

[59]  K. von Figura,et al.  A di‐leucine‐based motif in the cytoplasmic tail of LIMP‐II and tyrosinase mediates selective binding of AP‐3 , 1998, The EMBO journal.

[60]  Maria L. Wei,et al.  GLUT4 and Transferrin Receptor Are Differentially Sorted Along the Endocytic Pathway in CHO Cells , 1998, The Journal of cell biology.

[61]  S. Lemmon,et al.  The Yeast Adaptor Protein Complex, AP-3, Is Essential for the Efficient Delivery of Alkaline Phosphatase by the Alternate Pathway to the Vacuole , 1997, The Journal of cell biology.

[62]  Scott D Emr,et al.  The AP-3 Adaptor Complex Is Essential for Cargo-Selective Transport to the Yeast Vacuole , 1997, Cell.

[63]  T. Stevens,et al.  The Membrane Protein Alkaline Phosphatase Is Delivered to the Vacuole by a Route That Is Distinct from the VPS-dependent Pathway , 1997, The Journal of cell biology.

[64]  R. Kelly,et al.  ADP Ribosylation Factor 1 Is Required for Synaptic Vesicle Budding in PC12 Cells , 1997, The Journal of cell biology.

[65]  C. Geisler,et al.  Regulation and Function of the CD3γ DxxxLL Motif: A Binding Site for Adaptor Protein-1 and Adaptor Protein-2 in Vitro , 1997, The Journal of cell biology.

[66]  K. G. Coleman,et al.  Niemann-Pick C1 disease gene: homology to mediators of cholesterol homeostasis. , 1997, Science.

[67]  W. Pavan,et al.  Murine model of Niemann-Pick C disease: mutation in a cholesterol homeostasis gene. , 1997, Science.

[68]  J. Swedlow,et al.  Deconvolution in optical microscopy , 1996 .

[69]  R. Heilker,et al.  In vitro binding of clathrin adaptors to sorting signals correlates with endocytosis and basolateral sorting. , 1996, The EMBO journal.

[70]  K. von Figura,et al.  The Two Mannose 6-Phosphate Receptors Transport Distinct Complements of Lysosomal Proteins (*) , 1995, The Journal of Biological Chemistry.

[71]  M. R. Rad,et al.  Saccharomyces cerevisiae Apl2p, a homologue of the mammalian clathrin AP beta subunit, plays a role in clathrin-dependent Golgi functions. , 1995, Journal of cell science.

[72]  G. Payne,et al.  The Saccharomyces cerevisiae APS1 gene encodes a homolog of the small subunit of the mammalian clathrin AP‐1 complex: evidence for functional interaction with clathrin at the Golgi complex. , 1994, The EMBO journal.

[73]  M. Nakai,et al.  Cloning of the YAP19 gene encoding a putative yeast homolog of AP19, the mammalian small chain of the clathrin-assembly proteins. , 1993, Biochimica et biophysica acta.

[74]  T. Stevens,et al.  An MBoC Favorite: Morphological classification of the yeast vacuolar protein-sorting mutants: evidence for a prevacuolar compartment in class E vps mutants , 1992, Molecular biology of the cell.

[75]  S. Kornfeld,et al.  The cytoplasmic tail of the mannose 6-phosphate/insulin-like growth factor-II receptor has two signals for lysosomal enzyme sorting in the Golgi , 1992, The Journal of cell biology.

[76]  S. Kornfeld,et al.  A His-Leu-Leu sequence near the carboxyl terminus of the cytoplasmic domain of the cation-dependent mannose 6-phosphate receptor is necessary for the lysosomal enzyme sorting function. , 1992, The Journal of biological chemistry.

[77]  R. Klausner,et al.  A novel di-leucine motif and a tyrosine-based motif independently mediate lysosomal targeting and endocytosis of CD3 chains , 1992, Cell.

[78]  F. Maxfield,et al.  Iterative fractionation of recycling receptors from lysosomally destined ligands in an early sorting endosome , 1989, The Journal of cell biology.

[79]  L. Liscum,et al.  The intracellular transport of low density lipoprotein-derived cholesterol is defective in Niemann-Pick type C fibroblasts , 1989, The Journal of cell biology.

[80]  R. Brady,et al.  A defect in cholesterol esterification in Niemann-Pick disease (type C) patients. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[81]  A. Helenius,et al.  Glycoproteins of the lysosomal membrane , 1985, The Journal of cell biology.

[82]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[83]  Demel Ra,et al.  PENETRATION OF LIPID MONOLAYERS BY POLYENE ANTIBIOTICS. CORRELATION WITH SELECTIVE TOXICITY AND MODE OF ACTION. , 1965 .

[84]  P. De Camilli,et al.  Phosphatidylinositol-4-kinase type II alpha is a component of adaptor protein-3-derived vesicles. , 2005, Molecular biology of the cell.

[85]  K. Gernert,et al.  Saccharomyces cerevisiae Npc 2 p Is a Functionally Conserved Homologue of the Human Niemann-Pick Disease Type C 2 Protein , hNPC 2 † , 2005 .

[86]  J. Bonifacino,et al.  Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta 3A subunit of the AP-3 adaptor. , 1999, Molecular cell.

[87]  R. Kelly,et al.  ADP Ribosylation Factor 1 Is Required for Synaptic Vesicle Budding in PC 12 Cells , 1997 .

[88]  P. Jansson.  Deconvolution of images and spectra , 1997 .

[89]  T. Kirchhausen,et al.  A late Golgi sorting function for Saccharomyces cerevisiae Apm1p, but not for Apm2p, a second yeast clathrin AP medium chain-related protein. , 1995, Molecular biology of the cell.

[90]  I. Mellman,et al.  The biogenesis of lysosomes. , 1989, Annual review of cell biology.

[91]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[92]  R. Demel,et al.  PENETRATION OF LIPID MONOLAYERS BY POLYENE ANTIBIOTICS. CORRELATION WITH SELECTIVE TOXICITY AND MODE OF ACTION. , 1965, The Journal of biological chemistry.