In Polarized MDCK Cells Basolateral Vesicles Arise from Clathrin-γ-adaptin–coated Domains on Endosomal Tubules

Human transferrin receptors (TR) and receptors for polymeric immunoglobulins (pIgR) expressed in polarized MDCK cells maintain steady-state, asymmetric distributions on the separate basolateral and apical surfaces even though they are trafficking continuously into and across these cells. The intracellular mechanisms required to maintain these asymmetric distributions have not been located. Here we show that TR and pIgR internalize from both surfaces to a common interconnected endosome compartment that includes tubules with buds coated with clathrin lattices. These buds generate vesicles that carry TR to the basolateral border. The lattices contain γ-adaptin and are dispersed by treatment with brefeldin A (BFA). Since BFA treatment abrogates the vectorial trafficking of TR in polarized MDCK cells, we propose that the clathrin-coated domains of the endosome tubules contain the polarized sorting mechanism responsible for their preferential basolateral distribution.

[1]  I. Mellman,et al.  Inhibition of Endosome Function in CHO Cells Bearing a Temperature-sensitive Defect in the Coatomer (COPI) Component ε-COP , 1997, The Journal of cell biology.

[2]  F. Gu,et al.  Functional Dissection of COP-I Subunits in the Biogenesis of Multivesicular Endosomes , 1997, The Journal of cell biology.

[3]  I. Trowbridge,et al.  Structural Requirements for Basolateral Sorting of the Human Transferrin Receptor in the Biosynthetic and Endocytic Pathways of Madin-Darby Canine Kidney Cells , 1997, The Journal of cell biology.

[4]  T. Weimbs,et al.  Differential localization of syntaxin isoforms in polarized Madin-Darby canine kidney cells. , 1996, Molecular biology of the cell.

[5]  J. Bonifacino,et al.  Structural Determinants of Interaction of Tyrosine-based Sorting Signals with the Adaptor Medium Chains* , 1996, The Journal of Biological Chemistry.

[6]  C. Hopkins,et al.  Apical and basolateral endosomes of MDCK cells are interconnected and contain a polarized sorting mechanism , 1996, The Journal of cell biology.

[7]  W. Balch,et al.  Principles of selective transport: coat complexes hold the key. , 1996, Trends in cell biology.

[8]  G. Feldmann,et al.  The transcytotic pathway of an apical plasma membrane protein (B10) in hepatocytes is similar to that of IgA and occurs via a tubular pericentriolar compartment. , 1996, Journal of cell science.

[9]  J. Rothman,et al.  Protein Sorting by Transport Vesicles , 1996, Science.

[10]  F. Gu,et al.  An endosomal beta COP is involved in the pH-dependent formation of transport vesicles destined for late endosomes , 1996, The Journal of cell biology.

[11]  R. Schekman,et al.  Coat Proteins and Vesicle Budding , 1996, Science.

[12]  C. Futter,et al.  Multivesicular endosomes containing internalized EGF-EGF receptor complexes mature and then fuse directly with lysosomes , 1996, The Journal of cell biology.

[13]  B. Hoflack,et al.  Mannose 6-Phosphate Receptors and ADP-ribosylation Factors Cooperate for High Affinity Interaction of the AP-1 Golgi Assembly Proteins with Membranes (*) , 1996, The Journal of Biological Chemistry.

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

[15]  H. Nomoto,et al.  Anterograde and retrograde traffic between the rough endoplasmic reticulum and the Golgi complex , 1995, The Journal of cell biology.

[16]  I. Mellman,et al.  Cytoplasmic coat proteins involved in endosome function , 1995, Cell.

[17]  M. Spiess,et al.  Biosynthetic transport of the asialoglycoprotein receptor H1 to the cell surface occurs via endosomes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[18]  H. Hauri,et al.  Selective reentry of recycling cell surface glycoproteins to the biosynthetic pathway in human hepatocarcinoma HepG2 cells , 1995, The Journal of cell biology.

[19]  E. Ikonen,et al.  Different requirements for NSF, SNAP, and Rab proteins in apical and basolateral transport in MDCK cells , 1995, Cell.

[20]  C. Futter,et al.  Newly Synthesized Transferrin Receptors Can Be Detected in the Endosome before They Appear on the Cell Surface (*) , 1995, The Journal of Biological Chemistry.

[21]  C. Futter,et al.  Transport into and out of the Golgi complex studied by transfecting cells with cDNAs encoding horseradish peroxidase , 1994, The Journal of cell biology.

[22]  Barbara J. Reaves,et al.  Overexpression of TGN38/41 leads to mislocalisation of γ‐adaptin , 1994, FEBS Letters.

[23]  M. Krieger,et al.  Isolation of three classes of conditional lethal Chinese hamster ovary cell mutants with temperature-dependent defects in low density lipoprotein receptor stability and intracellular membrane transport. , 1994, The Journal of biological chemistry.

[24]  I. Mellman,et al.  Mechanisms of cell polarity: sorting and transport in epithelial cells. , 1994, Current opinion in cell biology.

[25]  R. Pepperkok,et al.  Coat proteins in intracellular membrane transport. , 1994, Current opinion in cell biology.

[26]  M. Krieger,et al.  Disruptions in Golgi structure and membrane traffic in a conditional lethal mammalian cell mutant are corrected by epsilon-COP , 1994, The Journal of cell biology.

[27]  K. Mostov,et al.  Polarized sorting of the polymeric immunoglobulin receptor in the exocytotic and endocytotic pathways is controlled by the same amino acids. , 1994, The EMBO journal.

[28]  G. Warren,et al.  The TGN38 glycoprotein contains two non-overlapping signals that mediate localization to the trans-Golgi network , 1994, The Journal of cell biology.

[29]  W. Balch,et al.  Rab1 and Ca2+ are required for the fusion of carrier vesicles mediating endoplasmic reticulum to Golgi transport , 1994, The Journal of cell biology.

[30]  S. Mayor,et al.  Brefeldin A causes structural and functional alterations of the trans-Golgi network of MDCK cells. , 1994, Journal of cell science.

[31]  K. Mostov,et al.  Receptor-mediated transcytosis of IgA in MDCK cells is via apical recycling endosomes , 1994, The Journal of cell biology.

[32]  E. Sztul,et al.  Basolateral to apical transcytosis in polarized cells is indirect and involves BFA and trimeric G protein sensitive passage through the apical endosome , 1994, The Journal of cell biology.

[33]  S. Wong,et al.  The SXYQRL sequence in the cytoplasmic domain of TGN38 plays a major role in trans-Golgi network localization. , 1993, The Journal of biological chemistry.

[34]  I. Mellman,et al.  Common signals control low density lipoprotein receptor sorting in endosomes and the Golgi complex of MDCK cells , 1993, Cell.

[35]  H. Varmus,et al.  A receptor for subgroup A Rous sarcoma virus is related to the low density lipoprotein receptor , 1993, Cell.

[36]  K. Willison,et al.  A 102 kDa subunit of a Golgi‐associated particle has homology to beta subunits of trimeric G proteins. , 1993, The EMBO journal.

[37]  F. Brodsky,et al.  Adaptor self‐aggregation, adaptor‐receptor recognition and binding of alpha‐adaptin subunits to the plasma membrane contribute to recruitment of adaptor (AP2) components of clathrin‐coated pits. , 1993, The EMBO journal.

[38]  L. Kühn,et al.  The internalization signal and the phosphorylation site of transferrin receptor are distinct from the main basolateral sorting information. , 1993, The EMBO journal.

[39]  Paul Tempst,et al.  SNAP receptors implicated in vesicle targeting and fusion , 1993, Nature.

[40]  J. Bonifacino,et al.  Localization of TGN38 to the trans-Golgi network: involvement of a cytoplasmic tyrosine-containing sequence , 1993, The Journal of cell biology.

[41]  S. White,et al.  Monoclonal antibodies against defined epitopes of the human transferrin receptor cytoplasmic tail. , 1992, Biochimica et biophysica acta.

[42]  M. Taub,et al.  Brefeldin A enhances receptor-mediated transcytosis of transferrin in filter-grown Madin-Darby canine kidney cells. , 1992, The Journal of biological chemistry.

[43]  F. Brodsky,et al.  100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties , 1992, The Journal of cell biology.

[44]  T. Kreis,et al.  Recruitment of coat proteins onto Golgi membranes in intact and permeabilized cells: Effects of brefeldin A and G protein activators , 1992, Cell.

[45]  R. Kelly,et al.  Low density lipoprotein receptor and cation-independent mannose 6- phosphate receptor are transported from the cell surface to the Golgi apparatus at equal rates in PC12 cells , 1992, The Journal of cell biology.

[46]  K. Mostov,et al.  Plasma membrane protein sorting in polarized epithelial cells , 1992, The Journal of cell biology.

[47]  B. Hoflack,et al.  Distribution of newly synthesized lysosomal enzymes in the endocytic pathway of normal rat kidney cells , 1991, The Journal of cell biology.

[48]  I. Mellman,et al.  Selective inhibition of transcytosis by brefeldin A in MDCK cells , 1991, Cell.

[49]  W. Brown,et al.  Brefeldin A causes a microtubule-mediated fusion of the trans-Golgi network and early endosomes , 1991, Cell.

[50]  J. Lippincott-Schwartz,et al.  Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic , 1991, Cell.

[51]  C. Hopkins,et al.  Transferrin receptors promote the formation of clathrin lattices , 1991, Cell.

[52]  M. Robinson,et al.  Cloning and expression of gamma-adaptin, a component of clathrin-coated vesicles associated with the Golgi apparatus , 1990, The Journal of cell biology.

[53]  J. Lippincott-Schwartz,et al.  Dissociation of a 110-kD peripheral membrane protein from the Golgi apparatus is an early event in brefeldin A action , 1990, The Journal of cell biology.

[54]  A. Wandinger-Ness,et al.  Polarized sorting in epithelia , 1990, Cell.

[55]  J. Lippincott-Schwartz,et al.  Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin a suggests an ER recycling pathway , 1990, Cell.

[56]  C. Hopkins,et al.  Role of the human transferrin receptor cytoplasmic domain in endocytosis: localization of a specific signal sequence for internalization , 1990, The Journal of cell biology.

[57]  H. Geuze,et al.  The pathways of endocytosed transferrin and secretory protein are connected in the trans-Golgi reticulum , 1988, The Journal of cell biology.

[58]  J. Rothman,et al.  A new type of coated vesicular carrier that appears not to contain clathrin: Its possible role in protein transport within the Golgi stack , 1986, Cell.

[59]  J. Slot,et al.  A new method of preparing gold probes for multiple-labeling cytochemistry. , 1985, European journal of cell biology.

[60]  M. Snider,et al.  Intracellular movement of cell surface receptors after endocytosis: resialylation of asialo-transferrin receptor in human erythroleukemia cells , 1985, The Journal of cell biology.

[61]  I. Trowbridge,et al.  Recombinant Rous sarcoma virus vectors for avian cells. , 1994, Methods in cell biology.

[62]  C. Hopkins,et al.  Signal-dependent membrane protein trafficking in the endocytic pathway. , 1993, Annual review of cell biology.

[63]  E. Rodriguez-Boulan,et al.  Polarity of epithelial and neuronal cells. , 1992, Annual review of cell biology.

[64]  M. Robinson,et al.  Clathrin, adaptors, and sorting. , 1990, Annual review of cell biology.

[65]  E. Weibel.  Stereological Methods. Practical methods for biological morphometry , 1979 .