Volume 272, Number 49, Issue of December 5, 1997 pp. 30748-30752
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Multiple Sorting Signals Determine Apical Localization of a Nonglycosylated Integral Membrane Protein

(Received for publication, July 1, 1997, and in revised form, September 22, 1997)

From the Centro de Biología Molecular "Severo Ochoa," Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain

In polarized cells, newly synthesized proteins are sorted in the trans-Golgi network and from there delivered to either the apical or basolateral membranes. Madin-Darby canine kidney (MDCK) cells have been widely used as a model system to study sorting determinants to the apical and basolateral surfaces. Whereas sorting signals for basolateral transmembrane proteins seem to reside in their cytoplasmic domains, apical determinants appear to reside in the N-glycans of secretory proteins or in the glycolipid tails of glycosylphosphatidylinositol-linked proteins. We show in this study that a surface-expressed form of CD3-, a nonglycosylated type I membrane protein, is exclusively targeted to the apical membrane in MDCK cells by a glycolipid-independent transport pathway. Deletion of the cytoplasmic tail does not affect its distribution, whereas deletion of the transmembrane domain results in secretion from both surfaces although still predominantly through the apical membrane. The transmembrane domain of CD3- appended to rat growth hormone, a secretory protein that lacks apical and basolateral determinants, promotes basolateral localization of the chimeric protein. However, a growth hormone chimera containing both the transmembrane and cytoplasmic domains of CD3- resulted in localization to the apical and basolateral membranes. These results suggest there are multiple determinants in CD3- that affect its distribution in polarized MDCK cells. Whereas the transmembrane domain contains a basolateral determinant, the ectodomain and the cytoplasmic domain contain apical determinants.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. S. Chmelar and N. M. Nathanson Identification of a Novel Apical Sorting Motif and Mechanism of Targeting of the M2 Muscarinic Acetylcholine Receptor J. Biol. Chem., November 17, 2006; 281(46): 35381 - 35396. [Abstract] [Full Text] [PDF]

				B. A. Potter, R. P. Hughey, and O. A. Weisz Role of N- and O-glycans in polarized biosynthetic sorting Am J Physiol Cell Physiol, January 1, 2006; 290(1): C1 - C10. [Abstract] [Full Text] [PDF]

				C. He, M. Hobert, L. Friend, and C. Carlin The Epidermal Growth Factor Receptor Juxtamembrane Domain Has Multiple Basolateral Plasma Membrane Localization Determinants, Including a Dominant Signal with a Polyproline Core J. Biol. Chem., October 4, 2002; 277(41): 38284 - 38293. [Abstract] [Full Text] [PDF]

				V. S. Subramanian, J. S. Marchant, I. Parker, and H. M. Said Intracellular trafficking/membrane targeting of human reduced folate carrier expressed in Xenopus oocytes Am J Physiol Gastrointest Liver Physiol, December 1, 2001; 281(6): G1477 - G1486. [Abstract] [Full Text] [PDF]

				B. M. Tam, O. L. Moritz, L. B. Hurd, and D. S. Papermaster Identification of an Outer Segment Targeting Signal in the COOH Terminus of Rhodopsin Using Transgenic Xenopus laevis J. Cell Biol., December 18, 2000; 151(7): 1369 - 1380. [Abstract] [Full Text] [PDF]

				S. Barman and D. P. Nayak Analysis of the Transmembrane Domain of Influenza Virus Neuraminidase, a Type II Transmembrane Glycoprotein, for Apical Sorting and Raft Association J. Virol., July 15, 2000; 74(14): 6538 - 6545. [Abstract] [Full Text]

				Z. Karim-Jimenez, N. Hernando, J. Biber, and H. Murer Requirement of a leucine residue for (apical) membrane expression of type IIb NaPi cotransporters PNAS, March 14, 2000; 97(6): 2916 - 2921. [Abstract] [Full Text] [PDF]

				C. Lipardi, L. Nitsch, and C. Zurzolo 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, February 1, 2000; 11(2): 531 - 542. [Abstract] [Full Text]

				N. Meerson, V Bello, J. Delaunay, T. Slimane, D Delautier, C Lenoir, G Trugnan, and M Maurice Intracellular traffic of the ecto-nucleotide pyrophosphatase/phosphodiesterase NPP3 to the apical plasma membrane of MDCK and Caco-2 cells: apical targeting occurs in the absence of N-glycosylation J. Cell Sci., January 12, 2000; 113(23): 4193 - 4202. [Abstract] [PDF]

				K Prydz and K. Dalen Synthesis and sorting of proteoglycans J. Cell Sci., January 1, 2000; 113(2): 193 - 205. [Abstract] [PDF]

				J. H. Benting, A. G. Rietveld, and K. Simons N-Glycans Mediate the Apical Sorting of a GPI-anchored, Raft-associated Protein in Madin-Darby Canine Kidney Cells J. Cell Biol., July 26, 1999; 146(2): 313 - 320. [Abstract] [Full Text] [PDF]

				R. Puertollano, F. Martin-Belmonte, J. Millan, M. del Carmen de Marco, J. P. Albar, L. Kremer, and M. A. Alonso The MAL Proteolipid Is Necessary for Normal Apical Transport and Accurate Sorting of the Influenza Virus Hemagglutinin in Madin-Darby Canine Kidney Cells J. Cell Biol., April 5, 1999; 145(1): 141 - 151. [Abstract] [Full Text] [PDF]

				R. Jacob, U. Preuss, P. Panzer, M. Alfalah, S. Quack, M. G. Roth, H. Naim, and H. Y. Naim Hierarchy of Sorting Signals in Chimeras of Intestinal Lactase-Phlorizin Hydrolase and the Influenza Virus Hemagglutinin J. Biol. Chem., March 19, 1999; 274(12): 8061 - 8067. [Abstract] [Full Text] [PDF]

				E. Orzech, K. Schlessinger, A. Weiss, C. T. Okamoto, and B. Aroeti Interactions of the AP-1 Golgi Adaptor with the Polymeric Immunoglobulin Receptor and Their Possible Role in Mediating Brefeldin A-sensitive Basolateral Targeting from the trans-Golgi Network J. Biol. Chem., January 22, 1999; 274(4): 2201 - 2215. [Abstract] [Full Text] [PDF]

				X. Zheng, D. Lu, and J. E. Sadler Apical Sorting of Bovine Enteropeptidase Does Not Involve Detergent-resistant Association with Sphingolipid-Cholesterol Rafts J. Biol. Chem., January 15, 1999; 274(3): 1596 - 1605. [Abstract] [Full Text] [PDF]

				C. Saunders, J. R. Keefer, C. A. Bonner, and L. E. Limbird Targeting of G Protein-coupled Receptors to the Basolateral Surface of Polarized Renal Epithelial Cells Involves Multiple, Non-contiguous Structural Signals J. Biol. Chem., September 11, 1998; 273(37): 24196 - 24206. [Abstract] [Full Text] [PDF]

				F. Martin-Belmonte, M. A. Alonso, X. Zhang, and P. Arvan Thyroglobulin Is Selected as Luminal Protein Cargo for Apical Transport via Detergent-resistant Membranes in Epithelial Cells J. Biol. Chem., December 22, 2000; 275(52): 41074 - 41081. [Abstract] [Full Text] [PDF]

				R. Martinez-Maza, I. Poyatos, B. Lopez-Corcuera, E. Nunez, C. Gimenez, F. Zafra, and C. Aragon The Role of N-Glycosylation in Transport to the Plasma Membrane and Sorting of the Neuronal Glycine Transporter GLYT2 J. Biol. Chem., January 12, 2001; 276(3): 2168 - 2173. [Abstract] [Full Text] [PDF]

				F. Martin-Belmonte, P. Arvan, and M. A. Alonso MAL Mediates Apical Transport of Secretory Proteins in Polarized Epithelial Madin-Darby Canine Kidney Cells J. Biol. Chem., December 21, 2001; 276(52): 49337 - 49342. [Abstract] [Full Text] [PDF]