Journal of Cell Science, Vol 113, Issue 19 3387-3398, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Inducible expression of claudin-1-myc but not occludin-VSV-G results in aberrant tight junction strand formation in MDCK cells

KM McCarthy, SA Francis, JM McCormack, J Lai, RA Rogers, IB Skare, RD Lynch and EE Schneeberger
Department of Pathology, Massachusetts General Hospital, Boston, MA 02114, USA.

Occludin and 18 distinct members of the claudin family are tetra-span transmembrane proteins that are localized in cell-specific tight junctions (TJs). A previous study showed that expression of chick occludin in Madin-Darby canine kidney (MDCK) cells raised transepithelial electrical resistance (TER) and, paradoxically, increased mannitol flux. In the present study, we employed epitope tagged canine occludin expression, under the control of the tetracycline repressible transactivator, to determine the extent to which the unexpected parallel increase in TER and mannitol flux was related to a structural mismatch between avian and canine occludins, which are only 50% identical. To determine whether the paradoxical changes in permeability was specific to occludin, we assessed the effect of over-expressing epitope tagged murine claudin-1. Our data revealed that over-expression of either of the epitope tagged mammalian tight junction proteins increased TER, mannitol and FITC-dextran flux. We observed a 2- and up to 5.6-fold over-expression of occludin-VSV-G and claudin-1-myc, respectively, with no change in ZO-1, endogenous occludin or claudin-1 expression. Confocal microscopy revealed that occludin-VSV-G, claudin-1-myc and ZO-1 co-localized at the TJ. In addition, claudin-1-myc formed aberrant strands along the lateral cell surface without an underlying ZO-1 scaffold. In fracture labeled replicas these strands consisted of claudin-1-myc with little accompanying occludin. These observations suggest that in epithelial cells claudin-1 can assemble into TJ strands without the participation of either ZO-1 or occludin. The proximity of the myc tag to the COOH-terminal YV sequence of claudin-1 appeared to interfere with its interaction with ZO-1, since over-expression of non-tagged claudin-1 increased TER but had a minimal effect on solute flux and no aberrant strands formed. From our data we conclude that differences in structure between avian and mammalian occludin do not account for the observed paradoxical increase in mannitol flux. Levels of ZO-1 remained unchanged despite substantial increases in induced TJ integral protein expression, suggesting that an imbalance between levels of ZO-1 and occludin or claudin-1 leads to altered regulation of pores through which non-charged solute flux occurs. We suggest that ion and solute flux are differentially regulated at the TJ.

This article has been cited by other articles:

				P. V. Afonso, S. Ozden, M.-C. Prevost, C. Schmitt, D. Seilhean, B. Weksler, P.-O. Couraud, A. Gessain, I. A. Romero, and P.-E. Ceccaldi Human Blood-Brain Barrier Disruption by Retroviral-Infected Lymphocytes: Role of Myosin Light Chain Kinase in Endothelial Tight-Junction Disorganization J. Immunol., August 15, 2007; 179(4): 2576 - 2583. [Abstract] [Full Text] [PDF]

				S. Angelow, R. El-Husseini, S. A. Kanzawa, and A. S. L. Yu Renal localization and function of the tight junction protein, claudin-19 Am J Physiol Renal Physiol, July 1, 2007; 293(1): F166 - F177. [Abstract] [Full Text] [PDF]

				A. Ikari, S. Matsumoto, H. Harada, K. Takagi, H. Hayashi, Y. Suzuki, M. Degawa, and M. Miwa Phosphorylation of paracellin-1 at Ser217 by protein kinase A is essential for localization in tight junctions J. Cell Sci., May 1, 2006; 119(9): 1781 - 1789. [Abstract] [Full Text] [PDF]

				D. F. Balkovetz Claudins at the gate: determinants of renal epithelial tight junction paracellular permeability Am J Physiol Renal Physiol, March 1, 2006; 290(3): F572 - F579. [Abstract] [Full Text] [PDF]

				S. Angelow, K.-J. Kim, and A. S. L. Yu Claudin-8 modulates paracellular permeability to acidic and basic ions in MDCK II cells J. Physiol., February 15, 2006; 571(1): 15 - 26. [Abstract] [Full Text] [PDF]

				C. J. Watson, C. J. Hoare, D. R. Garrod, G. L. Carlson, and G. Warhurst Interferon-{gamma} selectively increases epithelial permeability to large molecules by activating different populations of paracellular pores J. Cell Sci., November 15, 2005; 118(22): 5221 - 5230. [Abstract] [Full Text] [PDF]

				M. D. Alexandre, Q. Lu, and Y.-H. Chen Overexpression of claudin-7 decreases the paracellular Cl- conductance and increases the paracellular Na+ conductance in LLC-PK1 cells J. Cell Sci., June 15, 2005; 118(12): 2683 - 2693. [Abstract] [Full Text] [PDF]

				S W C van Mil, R H J Houwen, and L W J Klomp Genetics of familial intrahepatic cholestasis syndromes J. Med. Genet., June 1, 2005; 42(6): 449 - 463. [Abstract] [Full Text] [PDF]

				K. S. Matlin Clues to occludin. Focus on "Knockdown of occludin expression leads to diverse phenotypic alterations in epithelial cells" Am J Physiol Cell Physiol, June 1, 2005; 288(6): C1191 - C1192. [Full Text] [PDF]

				A. S. L. Yu, K. M. McCarthy, S. A. Francis, J. M. McCormack, J. Lai, R. A. Rogers, R. D. Lynch, and E. E. Schneeberger Knockdown of occludin expression leads to diverse phenotypic alterations in epithelial cells Am J Physiol Cell Physiol, June 1, 2005; 288(6): C1231 - C1241. [Abstract] [Full Text] [PDF]

				C. M. Van Itallie, T. M. Gambling, J. L. Carson, and J. M. Anderson Palmitoylation of claudins is required for efficient tight-junction localization J. Cell Sci., April 1, 2005; 118(7): 1427 - 1436. [Abstract] [Full Text] [PDF]

				K. J. Roux, S. A. Amici, B. S. Fletcher, and L. Notterpek Modulation of Epithelial Morphology, Monolayer Permeability, and Cell Migration by Growth Arrest Specific 3/Peripheral Myelin Protein 22 Mol. Biol. Cell, March 1, 2005; 16(3): 1142 - 1151. [Abstract] [Full Text] [PDF]

				A. Ikari, N. Hirai, M. Shiroma, H. Harada, H. Sakai, H. Hayashi, Y. Suzuki, M. Degawa, and K. Takagi Association of Paracellin-1 with ZO-1 Augments the Reabsorption of Divalent Cations in Renal Epithelial Cells J. Biol. Chem., December 24, 2004; 279(52): 54826 - 54832. [Abstract] [Full Text] [PDF]

				C. M. Van Itallie and J. M. Anderson The Molecular Physiology of Tight Junction Pores Physiology, December 1, 2004; 19(6): 331 - 338. [Abstract] [Full Text] [PDF]

				H. Wen, D. D. Watry, M. C. G. Marcondes, and H. S. Fox Selective Decrease in Paracellular Conductance of Tight Junctions: Role of the First Extracellular Domain of Claudin-5 Mol. Cell. Biol., October 1, 2004; 24(19): 8408 - 8417. [Abstract] [Full Text] [PDF]

				P. Acharya, J. Beckel, W. G. Ruiz, E. Wang, R. Rojas, L. Birder, and G. Apodaca Distribution of the tight junction proteins ZO-1, occludin, and claudin-4, -8, and -12 in bladder epithelium Am J Physiol Renal Physiol, August 1, 2004; 287(2): F305 - F318. [Abstract] [Full Text] [PDF]

				G. Bazzoni and E. Dejana Endothelial Cell-to-Cell Junctions: Molecular Organization and Role in Vascular Homeostasis Physiol Rev, July 1, 2004; 84(3): 869 - 901. [Abstract] [Full Text] [PDF]

				C. Rahner, M. Fukuhara, S. Peng, S. Kojima, and L. J. Rizzolo The apical and basal environments of the retinal pigment epithelium regulate the maturation of tight junctions during development J. Cell Sci., July 1, 2004; 117(15): 3307 - 3318. [Abstract] [Full Text] [PDF]

				E. E. Schneeberger and R. D. Lynch The tight junction: a multifunctional complex Am J Physiol Cell Physiol, June 1, 2004; 286(6): C1213 - C1228. [Abstract] [Full Text] [PDF]

				A. B. Singh and R. C. Harris Epidermal Growth Factor Receptor Activation Differentially Regulates Claudin Expression and Enhances Transepithelial Resistance in Madin-Darby Canine Kidney Cells J. Biol. Chem., January 30, 2004; 279(5): 3543 - 3552. [Abstract] [Full Text] [PDF]

				C. M. Van Itallie and J. M. Anderson The Role of Claudins in Determining Paracellular Charge Selectivity Proceedings of the ATS, January 1, 2004; 1(1): 38 - 41. [Abstract] [Full Text] [PDF]

				C. M. Van Itallie, A. S. Fanning, and J. M. Anderson Reversal of charge selectivity in cation or anion-selective epithelial lines by expression of different claudins Am J Physiol Renal Physiol, December 1, 2003; 285(6): F1078 - F1084. [Abstract] [Full Text]

				J. Yoo, A. Nichols, J. Mammen, I. Calvo, J. C. Song, R. T. Worrell, K. Matlin, and J. B. Matthews Bryostatin-1 enhances barrier function in T84 epithelia through PKC-dependent regulation of tight junction proteins Am J Physiol Cell Physiol, August 1, 2003; 285(2): C300 - C309. [Abstract] [Full Text] [PDF]

				O. R. Colegio, C. V. Itallie, C. Rahner, and J. M. Anderson Claudin extracellular domains determine paracellular charge selectivity and resistance but not tight junction fibril architecture Am J Physiol Cell Physiol, June 1, 2003; 284(6): C1346 - C1354. [Abstract] [Full Text] [PDF]

				A. S. L. Yu, A. H. Enck, W. I. Lencer, and E. E. Schneeberger Claudin-8 Expression in Madin-Darby Canine Kidney Cells Augments the Paracellular Barrier to Cation Permeation J. Biol. Chem., May 2, 2003; 278(19): 17350 - 17359. [Abstract] [Full Text] [PDF]

				S. Amasheh, N. Meiri, A. H. Gitter, T. Schoneberg, J. Mankertz, J. D. Schulzke, and M. Fromm Claudin-2 expression induces cation-selective channels in tight junctions of epithelial cells J. Cell Sci., March 14, 2003; 115(24): 4969 - 4976. [Abstract] [Full Text] [PDF]

				S. Peng, C. Rahner, and L. J. Rizzolo Apical and Basal Regulation of the Permeability of the Retinal Pigment Epithelium Invest. Ophthalmol. Vis. Sci., February 1, 2003; 44(2): 808 - 817. [Abstract] [Full Text] [PDF]

				L. L. Mitic, V. M. Unger, and J. M. Anderson Expression, solubilization, and biochemical characterization of the tight junction transmembrane protein claudin-4 Protein Sci., February 1, 2003; 12(2): 218 - 227. [Abstract] [Full Text] [PDF]

				G. Ranaldi, G. Perozzi, A. Truong-Tran, P. Zalewski, and C. Murgia Intracellular distribution of labile Zn(II) and zinc transporter expression in kidney and MDCK cells Am J Physiol Renal Physiol, December 1, 2002; 283(6): F1365 - F1375. [Abstract] [Full Text] [PDF]

				C. Y. Cheng and D. D. Mruk Cell Junction Dynamics in the Testis: Sertoli-Germ Cell Interactions and Male Contraceptive Development Physiol Rev, October 1, 2002; 82(4): 825 - 874. [Abstract] [Full Text] [PDF]

				O. R. Colegio, C. M. Van Itallie, H. J. McCrea, C. Rahner, and J. M. Anderson Claudins create charge-selective channels in the paracellular pathway between epithelial cells Am J Physiol Cell Physiol, July 1, 2002; 283(1): C142 - C147. [Abstract] [Full Text] [PDF]

				A.-M. Marzesco, I. Dunia, R. Pandjaitan, M. Recouvreur, D. Dauzonne, E. L. Benedetti, D. Louvard, and A. Zahraoui The Small GTPase Rab13 Regulates Assembly of Functional Tight Junctions in Epithelial Cells Mol. Biol. Cell, June 1, 2002; 13(6): 1819 - 1831. [Abstract] [Full Text] [PDF]

				L. Notterpek, K. J. Roux, S. A. Amici, A. Yazdanpour, C. Rahner, and B. S. Fletcher Peripheral myelin protein 22 is a constituent of intercellular junctions in epithelia PNAS, November 15, 2001; (2001) 251548398. [Abstract] [Full Text] [PDF]

				J. M. Anderson Molecular Structure of Tight Junctions and Their Role in Epithelial Transport Physiology, June 1, 2001; 16(3): 126 - 130. [Abstract] [Full Text] [PDF]

				M. Furuse, K. Furuse, H. Sasaki, and S. Tsukita Conversion of Zonulae Occludentes from Tight to Leaky Strand Type by Introducing Claudin-2 into Madin-Darby Canine Kidney I Cells J. Cell Biol., April 9, 2001; 153(2): 263 - 272. [Abstract] [Full Text] [PDF]

				L. Notterpek, K. J. Roux, S. A. Amici, A. Yazdanpour, C. Rahner, and B. S. Fletcher Peripheral myelin protein 22 is a constituent of intercellular junctions in epithelia PNAS, December 4, 2001; 98(25): 14404 - 14409. [Abstract] [Full Text] [PDF]