The Journal of Cell Biology, Vol 115, 607-618, Copyright © 1991 by The Rockefeller University Press

ARTICLES

An internal deletion in the cytoplasmic tail reverses the apical localization of human NGF receptor in transfected MDCK cells

A Le Bivic, Y Sambuy, A Patzak, N Patil, M Chao and E Rodriguez-Boulan
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.

A cDNA encoding the full-length 75-kD human nerve growth factor receptor was transfected into MDCK cells and its product was found to be expressed predominantly (80%) on the apical membrane, as a result of vectorial targeting from an intracellular site. Apical hNGFR bound NGF with low affinity and internalized it inefficiently (6% of surface bound NGF per hour). Several mutant hNGFRs were analyzed, after transfection in MDCK cells, for polarized surface expression, ligand binding, and endocytosis. Deletionof juxta-membrane attachment sites for a cluster of O-linked sugars did not alter apical localization. A mutant receptor lacking the entire cytoplasmic tail (except for the five proximal amino acids) was also expressed on the apical membrane, suggesting that information for apical sorting was contained in the ectoplasmic or transmembrane domains. However, a 58 amino acid deletion in the hNGFR tail that moved a cytoplasmic tyrosine (Tyr 308) closer to the membrane into a more charged environment resulted in a basolateral distribution of the mutant receptor and reversed vectorial (basolateral) targeting. The basolateral mutant receptor also internalized 125I-NGF rapidly (90% of surface bound NGF per hour), exhibited a larger intracellular fraction and displayed a considerably shortened half-life (approximately 3 h). We suggest that hNGFR with the internal cytoplasmic deletion expresses a basolateral targeting signal, related to endocytic signals, that is dominant over apical targeting information in the ecto/transmembrane domains. These results apparently contradict a current model that postulates that basolateral targeting is a default mechanism.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Jespersen, T., Rasmussen, H. B., Grunnet, M., Jensen, H. S., Angelo, K., Dupuis, D. S., Vogel, L. K., Jorgensen, N. K., Klaerke, D. A., Olesen, S.-P. (2004). Basolateral localisation of KCNQ1 potassium channels in MDCK cells: molecular identification of an N-terminal targeting motif. J. Cell Sci. 117: 4517-4526 [Abstract] [Full Text]  
• Beau, I., Groyer-Picard, M.-T., Desroches, A., Condamine, E., Leprince, J., Tome, J.-P., Dessen, P., Vaudry, H., Misrahi, M. (2004). The Basolateral Sorting Signals of the Thyrotropin and Luteinizing Hormone Receptors: An Unusual Family of Signals Sharing an Unusual Distal Intracellular Localization, but Unrelated in Their Structures. Mol. Endocrinol. 18: 733-746 [Abstract] [Full Text]  
• Lemmers, C., Michel, D., Lane-Guermonprez, L., Delgrossi, M.-H., Medina, E., Arsanto, J.-P., Le Bivic, A. (2004). CRB3 Binds Directly to Par6 and Regulates the Morphogenesis of the Tight Junctions in Mammalian Epithelial Cells. Mol. Biol. Cell 15: 1324-1333 [Abstract] [Full Text]  
• Brady, A. E., Wang, Q., Colbran, R. J., Allen, P. B., Greengard, P., Limbird, L. E. (2003). Spinophilin Stabilizes Cell Surface Expression of {alpha}2B-Adrenergic Receptors. J. Biol. Chem. 278: 32405-32412 [Abstract] [Full Text]  
• Jullien, J., Guili, V., Reichardt, L. F., Rudkin, B. B. (2002). Molecular Kinetics of Nerve Growth Factor Receptor Trafficking and Activation. J. Biol. Chem. 277: 38700-38708 [Abstract] [Full Text]  
• Dillon, C., Creer, A., Kerr, K., Kumin, A., Dickson, C. (2002). Basolateral Targeting of ERBB2 Is Dependent on a Novel Bipartite Juxtamembrane Sorting Signal but Independent of the C-Terminal ERBIN-Binding Domain. Mol. Cell. Biol. 22: 6553-6563 [Abstract] [Full Text]  
• Lipardi, C., Ruggiano, G., Perrone, L., Paladino, S., Monlauzeur, L., Nitsch, L., Le Bivic, A., Zurzolo, C. (2002). Differential Recognition of a Tyrosine-Dependent Signal in the Basolateral and Endocytic Pathways of Thyroid Epithelial Cells. Endocrinology 143: 1291-1301 [Abstract] [Full Text]  
• Straight, S. W., Chen, L., Karnak, D., Margolis, B. (2001). Interaction with mLin-7 Alters the Targeting of Endocytosed Transmembrane Proteins in Mammalian Epithelial Cells. Mol. Biol. Cell 12: 1329-1340 [Abstract] [Full Text]  
• Lipardi, C., Nitsch, L., Zurzolo, C. (2000). Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting. Mol. Biol. Cell 11: 531-542 [Abstract] [Full Text]  
• Watson, R. T., Pessin, J. E. (2000). Functional Cooperation of Two Independent Targeting Domains in Syntaxin 6 Is Required for Its Efficient Localization in the trans-Golgi Network of 3T3L1 Adipocytes. J. Biol. Chem. 275: 1261-1268 [Abstract] [Full Text]  
• YEAMAN, C., GRINDSTAFF, K. K., NELSON, W. J. (1999). New Perspectives on Mechanisms Involved in Generating Epithelial Cell Polarity. Physiol. Rev. 79: 73-98 [Abstract] [Full Text]  
• Monlauzeur, L., Breuza, L., Le Bivic, A. (1998). Putative O-Glycosylation Sites and a Membrane Anchor Are Necessary for Apical Delivery of the Human Neurotrophin Receptor in Caco-2 Cells. J. Biol. Chem. 273: 30263-30270 [Abstract] [Full Text]  
• Tugizov, S., Maidji, E., Xiao, J., Zheng, Z., Pereira, L. (1998). Human Cytomegalovirus Glycoprotein B Contains Autonomous Determinants for Vectorial Targeting to Apical Membranes of Polarized Epithelial Cells. J. Virol. 72: 7374-7386 [Abstract] [Full Text]  
• Marmorstein, A. D., Gan, Y. C., Bonilha, V. L., Finnemann, S. C., Csaky, K. G., Rodriguez-Boulan, E. (1998). Apical Polarity of N-CAM and EMMPRIN in Retinal Pigment Epithelium Resulting from Suppression of Basolateral Signal Recognition. JCB 142: 697-710 [Abstract] [Full Text]  
• Beau, I., Groyer-Picard, M.-T., Le Bivic, A., Vannier, B., Loosfelt, H., Milgrom, E., Misrahi, M. (1998). The Basolateral Localization Signal of the Follicle-stimulating Hormone Receptor. J. Biol. Chem. 273: 18610-18616 [Abstract] [Full Text]  
• Jou, T.-S., Schneeberger, E. E., James Nelson, W. (1998). Structural and Functional Regulation of Tight Junctions by RhoA and Rac1 Small GTPases. JCB 142: 101-115 [Abstract] [Full Text]  
• Yeaman, C., Gall, A. H. L., Baldwin, A. N., Monlauzeur, L., Bivic, A. L., Rodriguez-Boulan, E. (1997). The O-glycosylated Stalk Domain Is Required for Apical Sorting of Neurotrophin Receptors in Polarized MDCK Cells. JCB 139: 929-940 [Abstract] [Full Text]  
• Huang, X. F., Compans, R. W., Chen, S., Lamb, R. A., Arvan, P. (1997). Polarized Apical Targeting Directed by the Signal/Anchor Region of Simian Virus 5 Hemagglutinin-Neuraminidase. J. Biol. Chem. 272: 27598-27604 [Abstract] [Full Text]  
• Lin, S., Naim, H. Y., Roth, M. G. (1997). Tyrosine-dependent Basolateral Sorting Signals Are Distinct from Tyrosine-dependent Internalization Signals. J. Biol. Chem. 272: 26300-26305 [Abstract] [Full Text]  
• Maisner, A., Zimmer, G., Liszewski, M. K., Lublin, D. M., Atkinson, J. P., Herrler, G. (1997). Membrane Cofactor Protein (CD46) Is a Basolateral Protein That Is Not Endocytosed. IMPORTANCE OF THE TETRAPEPTIDE FTSL AT THE CARBOXYL TERMINUS. J. Biol. Chem. 272: 20793-20799 [Abstract] [Full Text]  
• Perego, C., Bulbarelli, A., Longhi, R., Caimi, M., Villa, A., Caplan, M. J., Pietrini, G. (1997). Sorting of Two Polytopic Proteins, the gamma -Aminobutyric Acid and Betaine Transporters, in Polarized Epithelial Cells. J. Biol. Chem. 272: 6584-6592 [Abstract] [Full Text]  
• Beau, I., Misrahi, M., Gross, B., Vannier, B., Loosfelt, H., Hai, M. T. V., Pichon, C., Milgrom, E. (1997). Basolateral Localization and Transcytosis of Gonadotropin and Thyrotropin Receptors Expressed in Madin-Darby Canine Kidney Cells. J. Biol. Chem. 272: 5241-5248 [Abstract] [Full Text]  
• Ure, D. R., Campenot, R. B. (1997). Retrograde Transport and Steady-State Distribution of 125I-Nerve Growth Factor in Rat Sympathetic Neurons in Compartmented Cultures. J. Neurosci. 17: 1282-1290 [Abstract] [Full Text]  
• Le Gall, A. H., Powell, S. K., Yeaman, C. A., Rodriguez-Boulan, E. (1997). The Neural Cell Adhesion Molecule Expresses a Tyrosine-independent Basolateral Sorting Signal. J. Biol. Chem. 272: 4559-4567 [Abstract] [Full Text]  
• Simonsen, A, Stang, E, Bremnes, B, Roe, M, Prydz, K, Bakke, O (1997). Sorting of MHC class II molecules and the associated invariant chain (Ii) in polarized MDCK cells. J. Cell Sci. 110: 597-609 [Abstract]  
• Maisner, A., Liszewski, M. K., Atkinson, J. P., Schwartz-Albiez, R., Herrler, G. (1996). Two Different Cytoplasmic Tails Direct Isoforms of the Membrane Cofactor Protein (CD46) to the Basolateral Surface of Madin-Darby Canine Kidney Cells. J. Biol. Chem. 271: 18853-18858 [Abstract] [Full Text]  
• Sheikh, H., Isacke, C. M. (1996). A Di-hydrophobic Leu-Val Motif Regulates the Basolateral Localization of CD44 in Polarized Madin-Darby Canine Kidney Epithelial Cells. J. Biol. Chem. 271: 12185-12190 [Abstract] [Full Text]  
• Noel, J, Roux, D, Pouyssegur, J (1996). Differential localization of Na+/H+ exchanger isoforms (NHE1 and NHE3) in polarized epithelial cell lines. J. Cell Sci. 109: 929-939 [Abstract]  
• Arreaza, G., Brown, D. A. (1995). Sorting and Intracellular Trafficking of a Glycosylphosphatidylinositol-anchored Protein and Two Hybrid Transmembrane Proteins with the Same Ectodomain in Madin-Darby Canine Kidney Epithelial Cells. J. Biol. Chem. 270: 23641-23647 [Abstract] [Full Text]  
• Monlauzeur, L., Rajasekaran, A., Chao, M., Rodriguez-Boulan, E., Bivic, A. L. (1995). A Cytoplasmic Tyrosine Is Essential for the Basolateral Localization of Mutants of the Human Nerve Growth Factor Receptor in Madin- Darby Canine Kidney Cells. J. Biol. Chem. 270: 12219-12225 [Abstract] [Full Text]  
• Wolf, D. E., McKinnon, C. A., Daou, M.-C., Stephens, R. M., Kaplan, D. R., Ross, A. H. (1995). Interaction with TrkA Immobilizes gp75 in the High Affinity Nerve Growth Factor Receptor Complex. J. Biol. Chem. 270: 2133-2138 [Abstract] [Full Text]  
• Mellman, I. (1995). Molecular Sorting of Membrane Proteins in Polarized and Nonpolarized Cells. Cold Spring Harb Symp Quant Biol 60: 745-752 [Abstract]  
• Mays, R.W., Nelson, W.J., Marrs, J.A. (1995). Generation of Epithelial Cell Polarity: Roles for Protein Trafficking, Membrane-Cytoskeleton, and E-Cadherin-mediated Cell Adhesion. Cold Spring Harb Symp Quant Biol 60: 763-773 [Abstract]  
• Garcia, M, Mirre, C, Quaroni, A, Reggio, H, Le Bivic, A (1993). GPI-anchored proteins associate to form microdomains during their intracellular transport in Caco-2 cells. J. Cell Sci. 104: 1281-1290 [Abstract]  
• Nelson, W. (1992). Regulation of cell surface polarity from bacteria to mammals. Science 258: 948-955 [Abstract]  
• Bailey, C., Chen, M, Keller, F, Kandel, E. (1992). Serotonin-mediated endocytosis of apCAM: an early step of learning-related synaptic growth in Aplysia. Science 256: 645-649 [Abstract]  
• Nelson, W.J., Wilson, R., Wollner, D., Mays, R., McNeill, H., Siemers, K. (1992). Regulation of Epithelial Cell Polarity: A View from the Cell Surface. Cold Spring Harb Symp Quant Biol 57: 621-630 [Abstract]  
• Nadler, L. S., Kumar, G., Nathanson, N. M. (2001). Identification of a Basolateral Sorting Signal for the M3 Muscarinic Acetylcholine Receptor in Madin-Darby Canine Kidney Cells. J. Biol. Chem. 276: 10539-10547 [Abstract] [Full Text]  
• Jaulin-Bastard, F., Saito, H., Le Bivic, A., Ollendorff, V., Marchetto, S., Birnbaum, D., Borg, J.-P. (2001). The ERBB2/HER2 Receptor Differentially Interacts with ERBIN and PICK1 PSD-95/DLG/ZO-1 Domain Proteins. J. Biol. Chem. 276: 15256-15263 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents