Identification and characterization of proteins that interact with zonula occludens proteins

IDENTIFICATION AND CHARACTERIZATION OF PROTEINS THAT INTERACT WITH ZONULA OCCLUDENS PROTEINS P JAYA KAUSALYA INSTITUTE OF MOLECULAR AND CELL BIOLOGY NATIONAL UNIVERSITY OF SINGAPORE 2005 IDENTIFICATION AND CHARACTERIZATION OF PROTEINS THAT INTERACT WITH ZONULA OCCLUDENS PROTEINS P JAYA KAUSALYA (B.Sc. (Hons.), NUS) A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY INSTITUTE OF MOLECULAR AND CELL BIOLOGY NATIONAL UNIVERSITY OF SINGAPORE 2005 ACKNOWLEGMENTS This work would not be possible without the support of family and colleagues. I wish to thank my parents and brother for their love, understanding and encouragement. I wish to thank A/P Walter Hunziker for being my mentor and I am grateful for his guidance and patience. I also, wish to thank my supervisory committee members, Prof Hong Wan Jin and A/P Cai Ming Jie for their guidance. I thank Dr. Manuela Reichert for her help during the initial stages of my project and collaborators, Dr. Dominik Muller and co-workers and Dr. Michael Fromme and co-workers. I am grateful to Dr. K. Willecke and Dr. Gonzalez-Mariscal for plasmids. I thank past and present members of WH lab and other IMCB members, in particular Drs. Wong Siew Cheng, Joy Tan, Lu Lei and Yu Xianwen for useful discussions and support. Last but not least, I thank God for all the opportunities given to me. TABLE OF CONTENTS LIST OF FIGURES LIST OF TABLES ABBREVIATIONS 10 MEDICAL DEFINITIONS 13 SUMMARY 14 CHAPTER 1: INTRODUCTION . 15 1.1: TIGHT JUNCTIONS 18 1.1.1: Tight junction structure and morphology . 18 1.2: Functions of Tight Junctions . 20 1.3: Models of Tight Junctions 23 1.4: Protein components of TJ 25 1.4.1: TJ transmembrane proteins . 26 1.4.1.1: Occludin . 27 1.4.1.1.1: Occludin as a structural and functional component of TJ 27 1.4.1.1.2: Protein-protein interactions of occludin . 30 1.4.1.2: Claudins . 31 1.4.1.2.1: Claudins as structural and functional components of TJs 34 1.4.1.2.2: Model for claudins in ion- and solute permeability 38 1.4.1.3: Junction Adhesion Molecule (JAM) 39 1.4.2: Peripherally-associated scaffolding proteins 42 1.4.2.1: PDZ domains . 43 1.4.2.2: Mechanism of binding and specificity of PDZ domains . 44 1.4.2.3: Structure and function of PDZ domain 45 1.4.2.4: Roles of PDZ domains . 49 1.4.2.5: SH3 and GUK domains . 50 1.4.2.6: The ZO protein family . 51 1.4.3: Regulatory proteins . 56 1.4.4: Transcriptional and post-transcriptional regulators 57 1.5: Assembly of Tight Junctions . 58 1.6: Diseases linked to the TJ function . 61 1.6.1: Diseases associated with TJ peripheral proteins . 61 1.6.2: Diseases associated with TJ integral membrane proteins . 62 1.7: Model systems to study junction assembly and function 65 1.7.1: General techniques 65 1.7.1.1: Epithelial cells as a model cell system . 65 1.7.1.2: Permeable supports . 66 1.7.1.3: Immunofluorescence analysis for TJ protein localization . 68 1.7.2: Analysis of TJ function . 69 1.7.2.1: Analysis of fence function . 69 1.7.2.2: Analysis of gate function . 70 CHAPTER 2: Identification and characterization of proteins that interact with ZO-1 PDZ domains using yeast-two-hybrid 71 Section 2.1: Connexin45 directly binds to the PDZ domains of ZO-1 and localizes to the tight junction region in epithelial MDCK cells . 73 2.1: Gap Junction and Connexins . 73 Section 2.2: Results . 76 Section 2.2.1: The C-terminus of Cx45 interacts with the PDZ domains of ZO-1 and ZO-3 in a yeast two-hybrid assay . 76 2.2.2: Characterization of epithelial MDCK cells transfected with Cx45 cDNA. 78 2.2.3: Cx45 directly associates with ZO-1 in vivo 80 2.2.4: Cx45 co-localizes with ZO-1 in the tight junction region in polarized MDCK cells 81 Section 2.3: Discussion . 84 Section 2.4: Association of ARVCF with ZO-1 and ZO-2: binding to PDZ-domain proteins and cell-cell adhesion regulate plasma membrane and nuclear localization of ARVCF . 89 2.4.1: Cadherins and Catenins 89 2.4.2: The p120 family 93 2.4.2.1: ARVCF 94 2.5: Results 96 2.5.1: ARVCF interacts with ZO-1 and ZO-2 but not ZO-3 96 2.5.2: ARVCF and ZO-1 interact in vivo and partially co-localize in MDCK cells . 102 2.5.4: ARVCF can associate with E-cadherin via its armadillo domains or through binding to PDZ domains proteins . 111 2.5.5: Plasma membrane and nuclear localization of ARVCF require the interaction with PDZ domain proteins and are regulated by cell-cell adhesion . 116 2.5.6: ARVCF, ZO-1 and E-cadherin are recruited to sites of initial cell-cell contact . 118 2.5.7: The PDZ domains of ZO-2 mediate the efficient nuclear localization of ARVCF . 122 Section 2.6: Discussion . 126 CHAPTER 3: Characterization of Claudin-16/Paracellin-1 and mutants associated with familial hypomagnesaemia with hypercalciuria and nephrocalcinosis (FHHNC) 132 Section 3.1: Claudin-16 132 3.1.1: Magnesium Homeostasis and Reabsorption . 132 3.1.2: Claudin-16/Paracellin-1 and renal magnesium wasting disorder . 133 3.1.3: Magnesium Resorption Mechanism . 135 3.1.4: Familial hypomagnesaemia with hypercalciuria and nephrocalcinosis linked to mutations in CLDN16 gene 137 3.1.5: Objective of study . 140 Section 3.2: Results on CLDN16 and mutations . 141 3.2.1: Characterization of an antibody to the first extracellular loop of CLDN16 . 141 3.2.2: Cldn16 internalizes via clathrin-dependent pathway 142 3.2.3: Subcellular and surface expression of Cldn16 mutations . 146 3.2.4: Cldn16 mutants that fail reach cell surface localize to the ER and Golgi 148 3.2.5: Characterization of trafficking defects of Cldn16 mutants displaying a predominant intracellular steady-state localization 151 3.2.5: Cldn16 mutants retained in the ER are subject to proteosomal degradation . 156 3.2.6: T233R mutation disruption PDZ binding motif and is mistargeted to the lysosomes 161 3.2.7: Cldn16 mutants delivered to lysosomes use different routes . 166 3.3.8: Pharmacological chaperones rescue cell surface expression of several Cldn16 mutants . 168 3.3.9: Cldn16 mutants present in TJ are defective in paracellular Mg2+ transport . 171 3.3.10: Clinical phenotypes . 175 Section 3.4: Discussion on Cldn16 and mutations 176 Chapter 4: Concluding Remarks . 185 Chapter 5: Materials and Methods 188 5.1 Antibodies and Reagents . 188 5.2: Plasmids constructions . 189 5.2.1: Cloning of ZO and mutants 189 5.2.2: Cloning of Connexin 45 and mutant constructs 189 5.2.3: Cloning of ARVCF and mutant constructs . 189 5.2.4: Cloning of Cld16 and mutants . 190 5.3: Yeast Two-Hybrid Screen . 190 5.4: Cell Cuture and Transfection of cells 191 5.5: Co-immunoprecipitation assays . 192 5.5.1:Cx45 coimmunoprecipitations . 192 5.5.2: ARVCF coimmunoprecipitation . 192 5.5.3: Cldn16 coimmunoprecipitation 193 5.6: GST Fusion Purification . 193 5.7: Pull-down assay . 194 5.7.1: ARVCF GST pull down assays 194 5.7.2: Cldn16 pull down assays 194 5.8: Immunofluorescence Labeling 195 5.9: Calcium-switch and cell-cell contact detection protocol . 196 5.10: Blot overlay 196 5.11: Cytochalasin D treatment for ARVCF or mutant transfected cells . 196 5.12: Endocytosis/ Internalization of CLDN16 and FHHNC mutants . 197 5.13: 20˚C block and cyclohexamide experiments . 197 5.14: Nocodazole treatment 197 5.15: Pharmacological inhibition of endocytosis 198 5.16: Proteosomal Degradation Western Analysis . 198 References . 199 LIST OF FIGURES Fig. 1: Junctional complexes. Fig. 2: Structure of Tight junctions. Fig. 3: Transcellular and Paracellular pathways. Fig. 4: Barrier and Fence function of tight junctions. Fig. 5: Proposed protein and lipid models of TJs. Fig. 6: The composition of tight junctions. Fig. 7: Integral membrane proteins of tight junctions. Fig. 8: Schematic representation of the structure and the membrane topology of claudins. Fig. 9: Creating a paracellular seal. Fig.10: Proposed model of ion and size discriminations and flux of charged and noncharged molecules. Fig. 11: Interaction of two of the major protein complexes in the TJs. Fig. 12: The ZO protein family in TJ that are part of MAGUK family. Fig. 13: Structure of PDZ3 domain of PSD-95 with a peptide ligand. Fig. 14: Possible modes of interaction of PDZ containing proteins. Fig. 15: ZO-3 interacts with PATJ, which is part of the Crumbs-Pals-PATJ complex. Fig.16: Stages of cell-cell contact and polarization. Fig. 17: Permeable supports use in the study of polarity. Fig. 18: TJ Barrier analysis. Fig. 19: Gap junctions between neighboring cells where ions and cAMP molecules are transferred. Fig. 20: Gap Junction Structure. Fig. 21: Characterization of MDCK cells expressing wild type or mutant Cx45. Fig. 22: Cx45 directly interacts with ZO-1 via the C-terminal SWVI. Fig. 23: Cx45 co-localizes with ZO-1 to the tight junction region in MDCK cells. Fig. 24: Main steps involved in connexin synthesis, assembly and turnover. Fig. 25: Schematic diagram of cadherin molecule showing its functional domains. Fig. 26: The armadillo family. Fig. 27: Cadherin-catenin complexes. Fig. 28: Schematic diagram of the domain structure of ARVCF. Fig. 29: Schematic diagrams of ARVCF, ZO-1 and ZO-2. Fig. 30: Binding of ARVCF and ZO-1. Fig. 31: A. ARVCF co-precipitates with ZO-1 from transfected MDCK cells. B. Endogenous ARVCF and ZO-1 coprecipitate from MDCK cells. Fig. 32: Colocalization of ARVCF and ZO-1 in transfected MDCK cells. Fig. 33: ARVCF partially colocalizes with ZO-1 to a discrete region between the lateral plasma membrane and TJ of polarized MDCK cells. Fig. 34: Colocalization and coprecipitation of ARVCF and ZO-1 or E-cadherin in transfected MDCK cells treated with cytochalasin D. Fig. 35: Binding of ARVCF and E-cadherin. Fig. 36: Plasma membrane recruitment and nuclear localization of ARVCF require the PDZ-binding motif and are regulated by cell-cell adhesion using calcium switch assay. Fig. 37: I. Recruitment of ARVCF to sites of cell-cell contact in MCF7 cells. II. Effect of ZO-1 mutants on ARVCF localization in MCF7 cells. Fig. 38: Role of ZO-2 in nuclear localization of ARVCF. Fig. 39: Putative E-cadherin/ARVCF/ZO-1 associations and C-terminal PDZ binding motifs in members of the p120ctn protein family. Fig. 40: Magnesium resorption in nephron segments. Fig. 41: Sequence, structure and expression of claudin-16. Fig. 42: A schematic model of magnesium resorption in the cortical ascending limb (cTAL) of Loop of Henle. Fig. 43: Predicted topology of CLDN16 and the location of the different mutations reported in humans. Fig. 44: Clathrin mediated endocytosis of Cldn16. Fig. 45: Cell surface expression of Cldn16 mutants linked to FHHNC. Fig. 46: Steady-state localization of Cldn16 mutants to different subcellular organelles. Fig. 47: Characterization of intracellular trafficking defects of different Cldn16 mutants. Fig. 48: Colocalization of Cldn16 mutants with ubiquitin is increased in the presence of a proteasome inhibitor. Fig. 49: ER-retained Cldn16 mutants are subject to proteasomal degradation. Fig. 50: Binding of wild type and mutant CLDN16 to ZO-1. Fig. 51: Subcellular localization of wild type and mutant CLDN16. Fig. 52: Cldn16 mutants that localize to lysosomes follow different pathways. Fig. 53: Chemical chaperones rescue cell surface expression of several Cldn16 mutants. Fig. 54: TJ localization of Cldn16 mutants expressed on the cell surface. Fig. 55: Measurements of Mg2+ permeability (A) and transepithelial resistance (B). Fig. 56: Predicted topology of Cldn16 and location of the different mutations linked to FHHNC reported. Fig. 57: Multiple roles of ZO proteins in the different adhesion junctional complexes. LIST OF TABLES Table 1: Claudin gene family Table 2: PDZ domain classes and examples of PDZ ligands. Table 3: Summary on disease associated with TJ peripheral proteins. Table 4: Summary on diseases associated with TJ integral membrane proteins. Table 5: Interaction of the PDZ domains of ZO-1, ZO-2 and ZO-3 with a construct encoding a C-terminal region of Cx45 or a Cx45 mutant in which the SVWI amino acids encoding a putative PDZ-binding motif were mutated to alanine. Table 6: Interaction of the PDZ domains of ZO-1 and ZO-2 with a construct encoding the C-terminal region of ARVCF (amino acids 670-893) or a mutant thereof (ARVCF∆P) in which the SWV amino acids encoding a putative PDZ-binding motif were changed to alanines. Table 7: Summary of the clinical features of FHHNC. Table 8: Summary of steady-state localization and defects of Cldn16 mutants linked to FHHNNC ABBREVIATIONS Å: Angstrom AF-6: ALL-1 fusion partner at chromosome AJ: Adherens Junction ALLN: N-acetyl–leu-leu-norleucinal Ap: Apical ARVCF: Armadillo repeat in Velo cardio facial syndrome Arm: Armadillo ASIP: atypical PKC isotype specific interacting protein BBB: Blood brain barrier Bl: Basolateral Ca2+: calcium CAR: Coxsackie and adenovirus receptors CPE: Clostridum perfringens enterotoxin Cld or Cldn: Claudin CLDN16: claudin-16 cTALH: cortical segment of the thick ascending limb of Henle Cx: Connexin Dlg: Drosophila Disc-Large DHR: Disc-large homology regions EEA1: Early Endosomal marker EMT: Epithelial-mesenchymal transition FHHNC: Familial Hypomagnesaemia with hypercalciuria and nephrocalcinosis 10 Izumi,Y., Hirose,T., Tamai,Y., Hirai,S., Nagashima,Y., Fujimoto,T., Tabuse,Y., Kemphues,K.J., and Ohno,S. (1998). An atypical PKC directly associates and colocalizes at the epithelial tight junction with ASIP, a mammalian homologue of Caenorhabditis elegans polarity protein PAR-3. J Cell Biol 143, 95-106. Jaramillo,B.E., Ponce,A., Moreno,J., Betanzos,A., Huerta,M., Lopez-Bayghen,E., and Gonzalez-Mariscal,L. (2004). Characterization of the tight junction protein ZO-2 localized at the nucleus of epithelial cells. Exp Cell Res 297, 247-258. Jarosch,E., Geiss-Friedlander,R., Meusser,B., Walter,J., and Sommer,T. (2002). Protein dislocation from the endoplasmic reticulum--pulling out the suspect. Traffic 3, 530-536. Jesaitis,L.A. and Goodenough,D.A. (1994). Molecular characterization and tissue distribution of ZO-2, a tight junction protein homologous to ZO-1 and the Drosophila discs-large tumor suppressor protein. J Cell Biol 124, 949-961. Johnson, L (2005). Applications of imaging techniques to studies of epithelial tight junctions. Adv Drug Delivery Reviews 57, 111-121. Johnston,J.A., Ward,C.L., and Kopito,R.R. (1998). Aggresomes: a cellular response to misfolded proteins. J Cell Biol 143, 1883-1898. Kachar,B. and Reese,T.S. (1982). Evidence for the lipidic nature of tight junction strands. Nature 296, 464-466. Kameya,S., Miyagoe,Y., Nonaka,I., Ikemoto,T., Endo,M., Hanaoka,K., Nabeshima,Y., and Takeda,S. (1999). alpha1-syntrophin gene disruption results in the absence of neuronal-type nitric-oxide synthase at the sarcolemma but does not induce muscle degeneration. J Biol Chem 274, 2193-2200. Kan,F.W. (1993). Cytochemical evidence for the presence of phospholipids in epithelial tight junction strands. J Histochem. Cytochem. 41, 649-656. Kanter,H.L., Laing,J.G., Beyer,E.C., Green,K.G., and Saffitz,J.E. (1993). Multiple connexins colocalize in canine ventricular myocyte gap junctions. Circ. Res 73, 344-350. Katahira,J., Sugiyama,H., Inoue,N., Horiguchi,Y., Matsuda,M., and Sugimoto,N. (1997). Clostridium perfringens enterotoxin utilizes two structurally related membrane proteins as functional receptors in vivo. J Biol Chem 272, 26652-26658. Kaufmann,U., Zuppinger,C., Waibler,Z., Rudiger,M., Urbich,C., Martin,B., Jockusch,B.M., Eppenberger,H., and Starzinski-Powitz,A. (2000). The armadillo repeat region targets ARVCF to cadherin-based cellular junctions. J Cell Sci 113 ( Pt 22), 41214135. Kim,E., Naisbitt,S., Hsueh,Y.P., Rao,A., Rothschild,A., Craig,A.M., and Sheng,M. (1997). GKAP, a novel synaptic protein that interacts with the guanylate kinase-like 208 domain of the PSD-95/SAP90 family of channel clustering molecules. J Cell Biol 136, 669-678. Kim,E., Niethammer,M., Rothschild,A., Jan,Y.N., and Sheng,M. (1995). Clustering of Shaker-type K+ channels by interaction with a family of membrane-associated guanylate kinases. Nature 378, 85-88. Kojima,T., Kokai,Y., Chiba,H., Yamamoto,M., Mochizuki,Y., and Sawada,N. (2001). Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes. Exp Cell Res 263, 193-201. Kojima,T., Sawada,N., Chiba,H., Kokai,Y., Yamamoto,M., Urban,M., Lee,G.H., Hertzberg,E.L., Mochizuki,Y., and Spray,D.C. (1999). Induction of tight junctions in human connexin 32 (hCx32)-transfected mouse hepatocytes: connexin 32 interacts with occludin. Biochem Biophys Res Commun 266, 222-229. Kollmar,R., Nakamura,S.K., Kappler,J.A., and Hudspeth,A.J. (2001). Expression and phylogeny of claudins in vertebrate primordia. Proc Natl Acad Sci U S A 98, 1019610201. Kominsky,S.L., Argani,P., Korz,D., Evron,E., Raman,V., Garrett,E., Rein,A., Sauter,G., Kallioniemi,O.P., and Sukumar,S. (2003). Loss of the tight junction protein claudin-7 correlates with histological grade in both ductal carcinoma in situ and invasive ductal carcinoma of the breast. Oncogene 22, 2021-2033. Kornecki,E., Walkowiak,B., Naik,U.P., and Ehrlich,Y.H. (1990). Activation of human platelets by a stimulatory monoclonal antibody. J Biol Chem 265, 10042-10048. Kruse,K., Kracht,U., and Kruse,U. (1984). Reference values for urinary calcium excretion and screening for hypercalciuria in children and adolescents. Eur J Pediatr. 143, 25-31. Kubota,K., Furuse,M., Sasaki,H., Sonoda,N., Fujita,K., Nagafuchi,A., and Tsukita,S. (1999). Ca(2+)-independent cell-adhesion activity of claudins, a family of integral membrane proteins localized at tight junctions. Curr Biol 9, 1035-1038. Kumar,N.M. and Gilula,N.B. (1996). The gap junction communication channel. Cell 84, 381-388. Kuriyama,M., Harada,N., Kuroda,S., Yamamoto,T., Nakafuku,M., Iwamatsu,A., Yamamoto,D., Prasad,R., Croce,C., Canaani,E., and Kaibuchi,K. (1996). Identification of AF-6 and canoe as putative targets for Ras. J Biol Chem 271, 607-610. Laing,J.G., Chou,B.C., and Steinberg,T.H. (2005). ZO-1 alters the plasma membrane localization and function of Cx43 in osteoblastic cells. J Cell Sci. 209 Laing,J.G., Manley-Markowski,R.N., Koval,M., Civitelli,R., and Steinberg,T.H. (2001). Connexin45 interacts with zonula occludens-1 and connexin43 in osteoblastic cells. J Biol Chem 276, 23051-23055. Laird,D.W., Castillo,M., and Kasprzak,L. (1995). Gap junction turnover, intracellular trafficking, and phosphorylation of connexin43 in brefeldin A-treated rat mammary tumor cells. J Cell Biol 131, 1193-1203. Laura,R.P., Witt,A.S., Held,H.A., Gerstner,R., Deshayes,K., Koehler,M.F., Kosik,K.S., Sidhu,S.S., and Lasky,L.A. (2002). The Erbin PDZ domain binds with high affinity and specificity to the carboxyl termini of delta-catenin and ARVCF. J Biol Chem 277, 1290612914. Lawrence,D.W., Comerford,K.M., and Colgan,S.P. (2002). Role of VASP in reestablishment of epithelial tight junction assembly after Ca2+ switch. Am J Physiol Cell Physiol 282, C1235-C1245. Lee,S.S., Glaunsinger,B., Mantovani,F., Banks,L., and Javier,R.T. (2000). Multi-PDZ domain protein MUPP1 is a cellular target for both adenovirus E4-ORF1 and high-risk papillomavirus type 18 E6 oncoproteins. J Virol. 74, 9680-9693. Lemmers,C, Michel,D, Lane-Gumeronprez,L, Delgrossi,M, Medina,E, Arsanto,J and Le Bivic (2004). CRB3 binds directly to Par6 and regulates the morphogenesis of the tight junction in mammalian epithelial cells. Mol Biol Cell, 15, 1324-1333. Lelievre-Pegorier,M., Merlet-Benichou,C., Roinel,N., and de Rouffignac,C. (1983). Developmental pattern of water and electrolyte transport in rat superficial nephrons. Am J Physiol 245, F15-F21. Leroy,A., Lauwaet,T., De Bruyne,G., Cornelissen,M., and Mareel,M. (2000). Entamoeba histolytica disturbs the tight junction complex in human enteric T84 cell layers. FASEB J 14, 1139-1146. Li,X., Olson,C., Lu,S., and Nagy,J.I. (2004). Association of connexin36 with zonula occludens-1 in HeLa cells, betaTC-3 cells, pancreas, and adrenal gland. Histochem. Cell Biol 122, 485-498. Liu,Y., Nusrat,A., Schnell,F.J., Reaves,T.A., Walsh,S., Pochet,M., and Parkos,C.A. (2000). Human junction adhesion molecule regulates tight junction resealing in epithelia. J Cell Sci 113 ( Pt 13), 2363-2374. Loh,Y.H., Christoffels,A., Brenner,S., Hunziker,W., and Venkatesh,B. (2004). Extensive expansion of the claudin gene family in the teleost fish, Fugu rubripes. Genome Res 14, 1248-1257. Lu,Q., Paredes,M., Medina,M., Zhou,J., Cavallo,R., Peifer,M., Orecchio,L., and Kosik,K.S. (1999). delta-catenin, an adhesive junction-associated protein which promotes cell scattering. J Cell Biol 144, 519-532. 210 Maekawa,K., Imagawa,N., Naito,A., Harada,S., Yoshie,O., and Takagi,S. (1999). Association of protein-tyrosine phosphatase PTP-BAS with the transcription-factorinhibitory protein IkappaBalpha through interaction between the PDZ1 domain and ankyrin repeats. Biochem J 337 ( Pt 2), 179-184. Mancini,A., Koch,A., Stefan,M., Niemann,H., and Tamura,T. (2000). The direct association of the multiple PDZ domain containing proteins (MUPP-1) with the human cKit C-terminus is regulated by tyrosine kinase activity. FEBS Lett 482, 54-58. Mandai,K., Nakanishi,H., Satoh,A., Obaishi,H., Wada,M., Nishioka,H., Itoh,M., Mizoguchi,A., Aoki,T., Fujimoto,T., Matsuda,Y., Tsukita,S., and Takai,Y. (1997). Afadin: A novel actin filament-binding protein with one PDZ domain localized at cadherin-based cell-to-cell adherens junction. J Cell Biol 139, 517-528. Mankertz,J., Waller,J.S., Hillenbrand,B., Tavalali,S., Florian,P., Schoneberg,T., Fromm,M., and Schulzke,J.D. (2002). Gene expression of the tight junction protein occludin includes differential splicing and alternative promoter usage. Biochem Biophys Res Commun 298, 657-666. Mariner,D.J., Wang,J., and Reynolds,A.B. (2000). ARVCF localizes to the nucleus and adherens junction and is mutually exclusive with p120(ctn) in E-cadherin complexes. J Cell Sci 113 ( Pt 8), 1481-1490. Martin-Padura,I., Lostaglio,S., Schneemann,M., Williams,L., Romano,M., Fruscella,P., Panzeri,C., Stoppacciaro,A., Ruco,L., Villa,A., Simmons,D., and Dejana,E. (1998). Junctional adhesion molecule, a novel member of the immunoglobulin superfamily that distributes at intercellular junctions and modulates monocyte transmigration. J Cell Biol 142, 117-127. Matlin,K.S. and Simons,K. (1983). Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation. Cell 34, 233-243. Mattagajasingh,S.N., Huang,S.C., Hartenstein,J.S., Snyder,M., Marchesi,V.T., and Benz,E.J. (1999). A nonerythroid isoform of protein 4.1R interacts with the nuclear mitotic apparatus (NuMA) protein. J Cell Biol 145, 29-43. Matter, K and Balda, M.S (2003). Functional analysis of tight junction. Methods 30, 228234. Matter, K and Balda, M.S (2003). Signaling to and from tight junctions. Nat Rev Mol Cell Biol, 4, 225-236. McCarthy,K.M., Skare,I.B., Stankewich,M.C., Furuse,M., Tsukita,S., Rogers,R.A., Lynch,R.D., and Schneeberger,E.E. (1996). Occludin is a functional component of the tight junction. J Cell Sci 109 ( Pt 9), 2287-2298. 211 Michel,D, Arsanto,J, Massey-Harroche,D, Beclin,C, Wijnholds, J and Le Bivic,A (2005). PATJ connects and stabilizes apical and lateral components of tight junctions in human intestinal cells. J Cell Sci, 118, 4049-4057. Milks,L.C., Kumar,N.M., Houghten,R., Unwin,N., and Gilula,N.B. (1988). Topology of the 32-kd liver gap junction protein determined by site-directed antibody localizations. EMBO J 7, 2967-2975. Mitic,L.L. and Anderson,J.M. (1998). Molecular architecture of tight junctions. Annu Rev Physiol 60, 121-142. Mitic,L.L., Schneeberger,E.E., Fanning,A.S., and Anderson,J.M. (1999). Connexinoccludin chimeras containing the ZO-binding domain of occludin localize at MDCK tight junctions and NRK cell contacts. J Cell Biol 146, 683-693. Mohandas,T.K., Chen,X.N., Rowe,L.B., Birkenmeier,E.H., Fanning,A.S., Anderson,J.M., and Korenberg,J.R. (1995). Localization of the tight junction protein gene TJP1 to human chromosome 15q13, distal to the Prader-Willi/Angelman region, and to mouse chromosome 7. Genomics 30, 594-597. Morita,K., Furuse,M., Fujimoto,K., and Tsukita,S. (1999). Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands. Proc Natl Acad Sci U S A 96, 511-516. Moss,A.L. and Ward,W.F. (1991). Multiple pathways for ligand internalization in rat hepatocytes. I: Effects of anoxia, phenylarsine oxide and monensin. J Cell Physiol 149, 313-318. Muller,D., Kausalya,P.J., Claverie-Martin,F., Meij,I.C., Eggert,P., Garcia-Nieto,V., and Hunziker,W. (2003). A novel claudin 16 mutation associated with childhood hypercalciuria abolishes binding to ZO-1 and results in lysosomal mistargeting. Am J Hum Genet 73, 1293-1301. Muller,D., Marr,N., Ankermann,T., Eggert,P., and Deen,P.M. (2002). Desmopressin for nocturnal enuresis in nephrogenic diabetes insipidus. Lancet 359, 495-497. Muller,T., Choidas,A., Reichmann,E., and Ullrich,A. (1999). Phosphorylation and free pool of beta-catenin are regulated by tyrosine kinases and tyrosine phosphatases during epithelial cell migration. J Biol Chem 274, 10173-10183. Muresan,Z., Paul,D.L., and Goodenough,D.A. (2000). Occludin 1B, a variant of the tight junction protein occludin. Mol Biol Cell 11, 627-634. Musil,L.S. and Goodenough,D.A. (1991). Biochemical analysis of connexin43 intracellular transport, phosphorylation, and assembly into gap junctional plaques. J Cell Biol 115, 1357-1374. 212 Nabi,I.R., Le Bivic,A., Fambrough,D., and Rodriguez-Boulan,E. (1991). An endogenous MDCK lysosomal membrane glycoprotein is targeted basolaterally before delivery to lysosomes. J Cell Biol 115, 1573-1584. Nagai-Tamai,Y., Mizuno,K., Hirose,T., Suzuki,A., and Ohno,S. (2002). Regulated protein-protein interaction between aPKC and PAR-3 plays an essential role in the polarization of epithelial cells. Genes Cells 7, 1161-1171. Naik,U.P., Ehrlich,Y.H., and Kornecki,E. (1995). Mechanisms of platelet activation by a stimulatory antibody: cross-linking of a novel platelet receptor for monoclonal antibody F11 with the Fc gamma RII receptor. Biochem J 310 ( Pt 1), 155-162. Nakamura,T., Blechman,J., Tada,S., Rozovskaia,T., Itoyama,T., Bullrich,F., Mazo,A., Croce,C.M., Geiger,B., and Canaani,E. (2000). huASH1 protein, a putative transcription factor encoded by a human homologue of the Drosophila ash1 gene, localizes to both nuclei and cell-cell tight junctions. Proc Natl Acad Sci U S A 97, 7284-7289. Nasdala,I., Wolburg-Buchholz,K., Wolburg,H., Kuhn,A., Ebnet,K., Brachtendorf,G., Samulowitz,U., Kuster,B., Engelhardt,B., Vestweber,D., and Butz,S. (2002). A transmembrane tight junction protein selectively expressed on endothelial cells and platelets. J Biol Chem 277, 16294-16303. Navarro,C., Nola,S., Audebert,S., Santoni,M.J., Arsanto,J.P., Ginestier,C., Marchetto,S., Jacquemier,J., Isnardon,D., Le Bivic,A., Birnbaum,D., and Borg,J.P. (2005). Junctional recruitment of mammalian Scribble relies on E-cadherin engagement. Oncogene. Nielsen,P.A., Baruch,A., Shestopalov,V.I., Giepmans,B.N., Dunia,I., Benedetti,E.L., and Kumar,N.M. (2003). Lens connexins alpha3Cx46 and alpha8Cx50 interact with zonula occludens protein-1 (ZO-1). Mol Biol Cell 14, 2470-2481. Nielsen,P.A., Beahm,D.L., Giepmans,B.N., Baruch,A., Hall,J.E., and Kumar,N.M. (2002). Molecular cloning, functional expression, and tissue distribution of a novel human gap junction-forming protein, connexin-31.9. Interaction with zona occludens protein-1. J Biol Chem 277, 38272-38283. Nitta,T., Hata,M., Gotoh,S., Seo,Y., Sasaki,H., Hashimoto,N., Furuse,M., and Tsukita,S. (2003). Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice. J Cell Biol 161, 653-660. Nix,S.L., Chishti,A.H., Anderson,J.M., and Walther,Z. (2000). hCASK and hDlg associate in epithelia, and their src homology and guanylate kinase domains participate in both intramolecular and intermolecular interactions. J Biol Chem 275, 41192-41200. Nusrat,A., Chen,J.A., Foley,C.S., Liang,T.W., Tom,J., Cromwell,M., Quan,C., and Mrsny,R.J. (2000a). The coiled-coil domain of occludin can act to organize structural and functional elements of the epithelial tight junction. J Biol Chem 275, 29816-29822. 213 Nusrat,A., Parkos,C.A., Verkade,P., Foley,C.S., Liang,T.W., Innis-Whitehouse,W., Eastburn,K.K., and Madara,J.L. (2000b). Tight junctions are membrane microdomains. J Cell Sci 113 ( Pt 10), 1771-1781. Ohba,Y., Kitagawa,H., Kitoh,K., Sasaki,Y., Takami,M., Shinkai,Y., and Kunieda,T. (2000). A deletion of the paracellin-1 gene is responsible for renal tubular dysplasia in cattle. Genomics 68, 229-236. Ohkubo,T. and Ozawa,M. (1999). p120(ctn) binds to the membrane-proximal region of the E-cadherin cytoplasmic domain and is involved in modulation of adhesion activity. J Biol Chem 274, 21409-21415. Ozawa,M. and Kemler,R. (1998). The membrane-proximal region of the E-cadherin cytoplasmic domain prevents dimerization and negatively regulates adhesion activity. J Cell Biol 142, 1605-1613. Pinto,d.S. and Kachar,B. (1982). On tight-junction structure. Cell 28, 441-450. Polak-Charcon,S., Shoham,J., and Ben Shaul,Y. (1978). Junction formation in trypsinized cells of human adenocarcinoma cell line. Exp Cell Res 116, 1-13. Ponting,C.P., Phillips,C., Davies,K.E., and Blake,D.J. (1997). PDZ domains: targeting signalling molecules to sub-membranous sites. Bioessays 19, 469-479. Prasad,R., Gu,Y., Alder,H., Nakamura,T., Canaani,O., Saito,H., Huebner,K., Gale,R.P., Nowell,P.C., Kuriyama,K., and . (1993). Cloning of the ALL-1 fusion partner, the AF-6 gene, involved in acute myeloid leukemias with the t(6;11) chromosome translocation. Cancer Res 53, 5624-5628. Rajasekaran,A.K., Hojo,M., Huima,T., and Rodriguez-Boulan,E. (1996). Catenins and zonula occludens-1 form a complex during early stages in the assembly of tight junctions. J Cell Biol 132, 451-463. Reichert,M., Muller,T., and Hunziker,W. (2000). The PDZ domains of zonula occludens1 induce an epithelial to mesenchymal transition of Madin-Darby canine kidney I cells. Evidence for a role of beta-catenin/Tcf/Lef signaling. J Biol Chem 275, 9492-9500. Reiser,J., Kriz,W., Kretzler,M., and Mundel,P. (2000). The glomerular slit diaphragm is a modified adherens junction. J Am Soc. Nephrol. 11, 1-8. Rescigno,M., Urbano,M., Valzasina,B., Francolini,M., Rotta,G., Bonasio,R., Granucci,F., Kraehenbuhl,J.P., and Ricciardi-Castagnoli,P. (2001). Dendritic cells express tight junction proteins and penetrate gut epithelial monolayers to sample bacteria. Nat Immunol 2, 361-367. Reynolds,A.B., Daniel,J., McCrea,P.D., Wheelock,M.J., Wu,J., and Zhang,Z. (1994). Identification of a new catenin: the tyrosine kinase substrate p120cas associates with Ecadherin complexes. Mol Cell Biol 14, 8333-8342. 214 Reynolds,A.B., Daniel,J.M., Mo,Y.Y., Wu,J., and Zhang,Z. (1996). The novel catenin p120cas binds classical cadherins and induces an unusual morphological phenotype in NIH3T3 fibroblasts. Exp Cell Res 225, 328-337. Reynolds,A.B., Herbert,L., Cleveland,J.L., Berg,S.T., and Gaut,J.R. (1992). p120, a novel substrate of protein tyrosine kinase receptors and of p60v-src, is related to cadherin-binding factors beta-catenin, plakoglobin and armadillo. Oncogene 7, 24392445. Reynolds,A.B., Roesel,D.J., Kanner,S.B., and Parsons,J.T. (1989). Transformationspecific tyrosine phosphorylation of a novel cellular protein in chicken cells expressing oncogenic variants of the avian cellular src gene. Mol Cell Biol 9, 629-638. Rico,M., Mukherjee,A., Konieczkowski,M., Bruggeman,L.A., Miller,R.T., Khan,S., Schelling,J.R., and Sedor,J.R. (2005). WT1-Interacting Protein and ZO-1 Translocate into Podocyte Nuclei after Puromycin Aminonucleoside Treatment. Am J Physiol Renal Physiol. Roh,M.H., Fan,S., Liu,C.J., and Margolis,B. (2003). The Crumbs3-Pals1 complex participates in the establishment of polarity in mammalian epithelial cells. J Cell Sci 116, 2895-2906. Roh,M.H., Makarova,O., Liu,C.J., Shin,K., Lee,S., Laurinec,S., Goyal,M., Wiggins,R., and Margolis,B. (2002). The Maguk protein, Pals1, functions as an adapter, linking mammalian homologues of Crumbs and Discs Lost. J Cell Biol 157, 161-172. Rubenstein,R.C., Egan,M.E., and Zeitlin,P.L. (1997). In vitro pharmacologic restoration of CFTR-mediated chloride transport with sodium 4-phenylbutyrate in cystic fibrosis epithelial cells containing delta F508-CFTR. J Clin Invest 100, 2457-2465. Rubenstein,R.C. and Zeitlin,P.L. (1998). A pilot clinical trial of oral sodium 4phenylbutyrate (Buphenyl) in deltaF508-homozygous cystic fibrosis patients: partial restoration of nasal epithelial CFTR function. Am J Respir. Crit Care Med 157, 484-490. Rubenstein,R.C. and Zeitlin,P.L. (2000). Sodium 4-phenylbutyrate downregulates Hsc70: implications for intracellular trafficking of DeltaF508-CFTR. Am J Physiol Cell Physiol 278, C259-C267. Ryeom,S.W., Paul,D., and Goodenough,D.A. (2000). Truncation mutants of the tight junction protein ZO-1 disrupt corneal epithelial cell morphology. Mol Biol Cell 11, 1687-1696. Saitou,M., Fujimoto,K., Doi,Y., Itoh,M., Fujimoto,T., Furuse,M., Takano,H., Noda,T., and Tsukita,S. (1998). Occludin-deficient embryonic stem cells can differentiate into polarized epithelial cells bearing tight junctions. J Cell Biol 141, 397-408. 215 Saitou,M., Furuse,M., Sasaki,H., Schulzke,J.D., Fromm,M., Takano,H., Noda,T., and Tsukita,S. (2000). Complex phenotype of mice lacking occludin, a component of tight junction strands. Mol Biol Cell 11, 4131-4142. Sandvig,K., Olsnes,S., Petersen,O.W., and van Deurs,B. (1987). Acidification of the cytosol inhibits endocytosis from coated pits. J Cell Biol 105, 679-689. Saras,J. and Heldin,C.H. (1996). PDZ domains bind carboxy-terminal sequences of target proteins. Trends Biochem Sci 21, 455-458. Sasaki,H., Matsui,C., Furuse,K., Mimori-Kiyosue,Y., Furuse,M., and Tsukita,S. (2003). Dynamic behavior of paired claudin strands within apposing plasma membranes. Proc Natl Acad Sci U S A 100, 3971-3976. Sato,S., Ward,C.L., Krouse,M.E., Wine,J.J., and Kopito,R.R. (1996). Glycerol reverses the misfolding phenotype of the most common cystic fibrosis mutation. J Biol Chem 271, 635-638. Scherer,S.S., Xu,Y.T., Nelles,E., Fischbeck,K., Willecke,K., and Bone,L.J. (1998). Connexin32-null mice develop demyelinating peripheral neuropathy. Glia 24, 8-20. Schluter,H., Wepf,R., Moll,I., and Franke,W.W. (2004). Sealing the live part of the skin: the integrated meshwork of desmosomes, tight junctions and curvilinear ridge structures in the cells of the uppermost granular layer of the human epidermis. Eur J Cell Biol 83, 655-665. Schmidt,A., Utepbergenov,D.I., Krause,G., and Blasig,I.E. (2001). Use of surface plasmon resonance for real-time analysis of the interaction of ZO-1 and occludin. Biochem Biophys Res Commun 288, 1194-1199. Schneeberger,E.E., Lynch,R.D., Kelly,C.A., and Rabito,C.A. (1988). Modulation of tight junction formation in clone MDCK cells by fatty acid supplementation. Am J Physiol 254, C432-C440. Schulzke,J.D., Gitter,A.H., Mankertz,J., Spiegel,S., Seidler,U., Amasheh,S., Saitou,M., Tsukita,S., and Fromm,M. (2005). Epithelial transport and barrier function in occludindeficient mice. Biochim. Biophys Acta 1669, 34-42. Sheng,M. and Sala,C. (2001). PDZ domains and the organization of supramolecular complexes. Annu Rev Neurosci. 24, 1-29. Sheth,B., Fontaine,J.J., Ponza,E., McCallum,A., Page,A., Citi,S., Louvard,D., Zahraoui,A., and Fleming,T.P. (2000). Differentiation of the epithelial apical junctional complex during mouse preimplantation development: a role for rab13 in the early maturation of the tight junction. Mech Dev 97, 93-104. Shibamoto,S., Hayakawa,M., Takeuchi,K., Hori,T., Miyazawa,K., Kitamura,N., Johnson,K.R., Wheelock,M.J., Matsuyoshi,N., Takeichi,M., and . (1995). Association of 216 p120, a tyrosine kinase substrate, with E-cadherin/catenin complexes. J Cell Biol 128, 949-957. Short,D.B., Trotter,K.W., Reczek,D., Kreda,S.M., Bretscher,A., Boucher,R.C., Stutts,M.J., and Milgram,S.L. (1998). An apical PDZ protein anchors the cystic fibrosis transmembrane conductance regulator to the cytoskeleton. J Biol Chem 273, 1979719801. Shprintzen,R.J., Goldberg,R.B., Lewin,M.L., Sidoti,E.J., Berkman,M.D., Argamaso,R.V., and Young,D. (1978). A new syndrome involving cleft palate, cardiac anomalies, typical facies, and learning disabilities: velo-cardio-facial syndrome. Cleft Palate J 15, 56-62. Shprintzen,R.J., Goldberg,R.B., Young,D., and Wolford,L. (1981). The velo-cardiofacial syndrome: a clinical and genetic analysis. Pediatrics 67, 167-172. Shprintzen,R.J., Siegel-Sadewitz,V.L., Amato,J., and Goldberg,R.B. (1985). Retrospective diagnoses of previously missed syndromic disorders among 1,000 patients with cleft lip, cleft palate, or both. Birth Defects Orig. Artic. Ser. 21, 85-92. Shurety,W., Bright,N.A., and Luzio,J.P. (1996). The effects of cytochalasin D and phorbol myristate acetate on the apical endocytosis of ricin in polarised Caco-2 cells. J Cell Sci 109 ( Pt 12), 2927-2935. Siliciano,J.D. and Goodenough,D.A. (1988). Localization of the tight junction protein, ZO-1, is modulated by extracellular calcium and cell-cell contact in Madin-Darby canine kidney epithelial cells. J Cell Biol 107, 2389-2399. Simon,A.M., Goodenough,D.A., Li,E., and Paul,D.L. (1997). Female infertility in mice lacking connexin 37. Nature 385, 525-529. Simon,D.B., Lu,Y., Choate,K.A., Velazquez,H., Al Sabban,E., Praga,M., Casari,G., Bettinelli,A., Colussi,G., Rodriguez-Soriano,J., McCredie,D., Milford,D., Sanjad,S., and Lifton,R.P. (1999). Paracellin-1, a renal tight junction protein required for paracellular Mg2+ resorption. Science 285, 103-106. Simske,J.S., Koppen,M., Sims,P., Hodgkin,J., Yonkof,A., and Hardin,J. (2003). The cell junction protein VAB-9 regulates adhesion and epidermal morphology in C. elegans. Nat Cell Biol 5, 619-625. Singer,K.L., Stevenson,B.R., Woo,P.L., and Firestone,G.L. (1994). Relationship of serine/threonine phosphorylation/dephosphorylation signaling to glucocorticoid regulation of tight junction permeability and ZO-1 distribution in nontransformed mammary epithelial cells. J Biol Chem 269, 16108-16115. Sirotkin,H., O'Donnell,H., DasGupta,R., Halford,S., St Jore,B., Puech,A., Parimoo,S., Morrow,B., Skoultchi,A., Weissman,S.M., Scambler,P., and Kucherlapati,R. (1997). Identification of a new human catenin gene family member (ARVCF) from the region deleted in velo-cardio-facial syndrome. Genomics 41, 75-83. 217 Sitia,R. and Braakman,I. (2003). Quality control in the endoplasmic reticulum protein factory. Nature 426, 891-894. Smart,E.J., Ying,Y.S., Conrad,P.A., and Anderson,R.G. (1994). Caveolin moves from caveolae to the Golgi apparatus in response to cholesterol oxidation. J Cell Biol 127, 1185-1197. Sommers,C.L., Byers,S.W., Thompson,E.W., Torri,J.A., and Gelmann,E.P. (1994). Differentiation state and invasiveness of human breast cancer cell lines. Breast Cancer Res Treat. 31, 325-335. Songyang,Z., Fanning,A.S., Fu,C., Xu,J., Marfatia,S.M., Chishti,A.H., Crompton,A., Chan,A.C., Anderson,J.M., and Cantley,L.C. (1997). Recognition of unique carboxylterminal motifs by distinct PDZ domains. Science 275, 73-77. Sonoda,N., Furuse,M., Sasaki,H., Yonemura,S., Katahira,J., Horiguchi,Y., and Tsukita,S. (1999). Clostridium perfringens enterotoxin fragment removes specific claudins from tight junction strands: Evidence for direct involvement of claudins in tight junction barrier. J Cell Biol 147, 195-204. Staddon,J.M., Smales,C., Schulze,C., Esch,F.S., and Rubin,L.L. (1995). p120, a p120related protein (p100), and the cadherin/catenin complex. J Cell Biol 130, 369-381. Stankewich,M.C., Francis,S.A., Vu,Q.U., Schneeberger,E.E., and Lynch,R.D. (1996). Alterations in cell cholesterol content modulate Ca(2+)-induced tight junction assembly by MDCK cells. Lipids 31, 817-828. Stevenson,B.R., Anderson,J.M., Braun,I.D., and Mooseker,M.S. (1989). Phosphorylation of the tight-junction protein ZO-1 in two strains of Madin-Darby canine kidney cells which differ in transepithelial resistance. Biochem J 263, 597-599. Stevenson,B.R., Anderson,J.M., Goodenough,D.A., and Mooseker,M.S. (1988). Tight junction structure and ZO-1 content are identical in two strains of Madin-Darby canine kidney cells which differ in transepithelial resistance. J Cell Biol 107, 2401-2408. Stevenson,B.R. and Goodenough,D.A. (1984). Zonulae occludentes in junctional complex-enriched fractions from mouse liver: preliminary morphological and biochemical characterization. J Cell Biol 98, 1209-1221. Stevenson,B.R., Siliciano,J.D., Mooseker,M.S., and Goodenough,D.A. (1986). Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia. J Cell Biol 103, 755-766. Stuart,R.O. and Nigam,S.K. (1995). Regulated assembly of tight junctions by protein kinase C. Proc Natl Acad Sci U S A 92, 6072-6076. Sudol,M. (1998). From Src Homology domains to other signaling modules: proposal of the 'protein recognition code'. Oncogene 17, 1469-1474. 218 Suzuki,A., Ishiyama,C., Hashiba,K., Shimizu,M., Ebnet,K., and Ohno,S. (2002). aPKC kinase activity is required for the asymmetric differentiation of the premature junctional complex during epithelial cell polarization. J Cell Sci 115, 3565-3573. Suzuki,A., Yamanaka,T., Hirose,T., Manabe,N., Mizuno,K., Shimizu,M., Akimoto,K., Izumi,Y., Ohnishi,T., and Ohno,S. (2001). Atypical protein kinase C is involved in the evolutionarily conserved par protein complex and plays a critical role in establishing epithelia-specific junctional structures. J Cell Biol 152, 1183-1196. Takahashi,K., Nakanishi,H., Miyahara,M., Mandai,K., Satoh,K., Satoh,A., Nishioka,H., Aoki,J., Nomoto,A., Mizoguchi,A., and Takai,Y. (1999). Nectin/PRR: an immunoglobulin-like cell adhesion molecule recruited to cadherin-based adherens junctions through interaction with Afadin, a PDZ domain-containing protein. J Cell Biol 145, 539-549. Tamarappoo,B.K., Yang,B., and Verkman,A.S. (1999). Misfolding of mutant aquaporin2 water channels in nephrogenic diabetes insipidus. J Biol Chem 274, 34825-34831. Tamer,C.M., Lamm,M.E., Robinson,J.K., Piskurich,J.F., and Kaetzel,C.S. (1995). Comparative studies of transcytosis and assembly of secretory IgA in Madin-Darby canine kidney cells expressing human polymeric Ig receptor. J Immunol 155, 707-714. Tan,C.M., Nickols,H.H., and Limbird,L.E. (2003). Appropriate polarization following pharmacological rescue of V2 vasopressin receptors encoded by X-linked nephrogenic diabetes insipidus alleles involves a conformation of the receptor that also attains mature glycosylation. J Biol Chem 278, 35678-35686. Thomas,M., Glaunsinger,B., Pim,D., Javier,R., and Banks,L. (2001). HPV E6 and MAGUK protein interactions: determination of the molecular basis for specific protein recognition and degradation. Oncogene 20, 5431-5439. Thoreson,M.A., Anastasiadis,P.Z., Daniel,J.M., Ireton,R.C., Wheelock,M.J., Johnson,K.R., Hummingbird,D.K., and Reynolds,A.B. (2000). Selective uncoupling of p120(ctn) from E-cadherin disrupts strong adhesion. J Cell Biol 148, 189-202. Tobioka,H., Tokunaga,Y., Isomura,H., Kokai,Y., Yamaguchi,J., and Sawada,N. (2004). Expression of occludin, a tight-junction-associated protein, in human lung carcinomas. Virchows Arch. 445, 472-476. Toyofuku,T., Akamatsu,Y., Zhang,H., Kuzuya,T., Tada,M., and Hori,M. (2001). c-Src regulates the interaction between connexin-43 and ZO-1 in cardiac myocytes. J Biol Chem 276, 1780-1788. Toyofuku,T., Yabuki,M., Otsu,K., Kuzuya,T., Hori,M., and Tada,M. (1998). Direct association of the gap junction protein connexin-43 with ZO-1 in cardiac myocytes. J Biol Chem 273, 12725-12731. 219 Traweger,A., Fang,D., Liu,Y.C., Stelzhammer,W., Krizbai,I.A., Fresser,F., Bauer,H.C., and Bauer,H. (2002). The tight junction-specific protein occludin is a functional target of the E3 ubiquitin-protein ligase itch. J Biol Chem 277, 10201-10208. Tsukita,S., Furuse,M., and Itoh,M. (2001). Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2, 285-293. Tsukita, S, Furuse, M and Itoh, M (1999). Structural and signaling molecules come together at tight junction. Curr Opin in Cell Biol, 11, 628-633. Tsukita , S and Furuse, M (2000). Pores in the Wall: Claudins constitute tight junction strands containing aqueous pores. J Cell Biol, 149, 13-16. Tsunoda,S., Sierralta,J., Sun,Y., Bodner,R., Suzuki,E., Becker,A., Socolich,M., and Zuker,C.S. (1997). A multivalent PDZ-domain protein assembles signalling complexes in a G-protein-coupled cascade. Nature 388, 243-249. Turksen,K. and Troy,T.C. (2002). Permeability barrier dysfunction in transgenic mice overexpressing claudin 6. Development 129, 1775-1784. Turksen, K and Troy, T.C (2004). Barriers built on claudins. J Cell Sci, 117, 2435-2447. Ulloa-Aguirre,A., Janovick,J.A., Brothers,S.P., and Conn,P.M. (2004). Pharmacologic rescue of conformationally-defective proteins: implications for the treatment of human disease. Traffic 5, 821-837. Umeda,K., Matsui,T., Nakayama,M., Furuse,K., Sasaki,H., Furuse,M., and Tsukita,S. (2004). Establishment and characterization of cultured epithelial cells lacking expression of ZO-1. J Biol Chem 279, 44785-44794. Van Itallie,C., Rahner,C., and Anderson,J.M. (2001). Regulated expression of claudin-4 decreases paracellular conductance through a selective decrease in sodium permeability. J Clin Invest 107, 1319-1327. Van Itallie,C.M. and Anderson,J.M. (1997). Occludin confers adhesiveness when expressed in fibroblasts. J Cell Sci 110 ( Pt 9), 1113-1121. Van Itallie,C.M., Balda,M.S., and Anderson,J.M. (1995). Epidermal growth factor induces tyrosine phosphorylation and reorganization of the tight junction protein ZO-1 in A431 cells. J Cell Sci 108 ( Pt 4), 1735-1742. Van Itallie, C.M and Anderson, J.M (2004). The Role of Claudins in determining Paracellular Charge Selevtivity. Proc Am Thorac Soc 1, 38-41. Van Itallie, C.M, Fanning, A.S and Anderson, J.M (2005). Reversal of charge selectivity in cation or anion-selective epithelial lines by expression of different claudins. AJPRenal Physiol, 285, 1078-1084. 220 van Meer,G., Gumbiner,B., and Simons,K. (1986). The tight junction does not allow lipid molecules to diffuse from one epithelial cell to the next. Nature 322, 639-641. VanSlyke,J.K., Deschenes,S.M., and Musil,L.S. (2000). Intracellular transport, assembly, and degradation of wild-type and disease-linked mutant gap junction proteins. Mol Biol Cell 11, 1933-1946. Vleminckx,K. and Kemler,R. (1999). Cadherins and tissue formation: integrating adhesion and signaling. Bioessays 21, 211-220. Vincent,P.A, Xiao,K, Buckley,K.M and Kowalczyk, A.P (2004). VE-cadherin: adhesion at arm's length. AJP-Cell Physiol 286, C987-997. Waibler,Z., Schafer,A., and Starzinski-Powitz,A. (2001). mARVCF cellular localisation and binding to cadherins is influenced by the cellular context but not by alternative splicing. J Cell Sci 114, 3873-3884. Weber,E., Berta,G., Tousson,A., St John,P., Green,M.W., Gopalokrishnan,U., Jilling,T., Sorscher,E.J., Elton,T.S., Abrahamson,D.R., and . (1994). Expression and polarized targeting of a rab3 isoform in epithelial cells. J Cell Biol 125, 583-594. Weber,S., Hoffmann,K., Jeck,N., Saar,K., Boeswald,M., Kuwertz-Broeking,E., Meij,I.I., Knoers,N.V., Cochat,P., Sulakova,T., Bonzel,K.E., Soergel,M., Manz,F., Schaerer,K., Seyberth,H.W., Reis,A., and Konrad,M. (2000). Familial hypomagnesaemia with hypercalciuria and nephrocalcinosis maps to chromosome 3q27 and is associated with mutations in the PCLN-1 gene. Eur J Hum Genet 8, 414-422. Weber,S., Schlingmann,K.P., Peters,M., Nejsum,L.N., Nielsen,S., Engel,H., Grzeschik,K.H., Seyberth,H.W., Grone,H.J., Nusing,R., and Konrad,M. (2001a). Primary gene structure and expression studies of rodent paracellin-1. J Am Soc. Nephrol. 12, 2664-2672. Weber,S., Schneider,L., Peters,M., Misselwitz,J., Ronnefarth,G., Boswald,M., Bonzel,K.E., Seeman,T., Sulakova,T., Kuwertz-Broking,E., Gregoric,A., Palcoux,J.B., Tasic,V., Manz,F., Scharer,K., Seyberth,H.W., and Konrad,M. (2001b). Novel paracellin1 mutations in 25 families with familial hypomagnesemia with hypercalciuria and nephrocalcinosis. J Am Soc. Nephrol. 12, 1872-1881. Wick,W., Petersen,I., Schmutzler,R.K., Wolfarth,B., Lenartz,D., Bierhoff,E., Hummerich,J., Muller,D.J., Stangl,A.P., Schramm,J., Wiestler,O.D., and von Deimling,A. (1996). Evidence for a novel tumor suppressor gene on chromosome 15 associated with progression to a metastatic stage in breast cancer. Oncogene 12, 973978. Wilcox,E.R., Burton,Q.L., Naz,S., Riazuddin,S., Smith,T.N., Ploplis,B., Belyantseva,I., Ben Yosef,T., Liburd,N.A., Morell,R.J., Kachar,B., Wu,D.K., Griffith,A.J., Riazuddin,S., and Friedman,T.B. (2001). Mutations in the gene encoding tight junction claudin-14 cause autosomal recessive deafness DFNB29. Cell 104, 165-172. 221 Willecke,K., Eiberger,J., Degen,J., Eckardt,D., Romualdi,A., Guldenagel,M., Deutsch,U., and Sohl,G. (2002). Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem 383, 725-737. Williams,C.D. and Rizzolo,L.J. (1997). Remodeling of junctional complexes during the development of the outer blood-retinal barrier. Anat. Rec. 249, 380-388. Williams,M.A., Shprintzen,R.J., and Goldberg,R.B. (1985). Male-to-male transmission of the velo-cardio-facial syndrome: a case report and review of 60 cases. J Craniofac. Genet Dev Biol 5, 175-180. Wittchen,E.S., Haskins,J., and Stevenson,B.R. (1999). Protein interactions at the tight junction. Actin has multiple binding partners, and ZO-1 forms independent complexes with ZO-2 and ZO-3. J Biol Chem 274, 35179-35185. Wolf,M.T., Dotsch,J., Konrad,M., Boswald,M., and Rascher,W. (2002). Follow-up of five patients with FHHNC due to mutations in the Paracellin-1 gene. Pediatr. Nephrol. 17, 602-608. Woods,D.F. and Bryant,P.J. (1993). ZO-1, DlgA and PSD-95/SAP90: homologous proteins in tight, septate and synaptic cell junctions. Mech Dev 44, 85-89. Wu,V.M., Schulte,J., Hirschi,A., Tepass,U., and Beitel,G.J. (2004). Sinuous is a Drosophila claudin required for septate junction organization and epithelial tube size control. J Cell Biol 164, 313-323. Xia,H., Winokur,S.T., Kuo,W.L., Altherr,M.R., and Bredt,D.S. (1997). Actininassociated LIM protein: identification of a domain interaction between PDZ and spectrin-like repeat motifs. J Cell Biol 139, 507-515. Xu,X.Z., Choudhury,A., Li,X., and Montell,C. (1998). Coordination of an array of signaling proteins through homo- and heteromeric interactions between PDZ domains and target proteins. J Cell Biol 142, 545-555. Yamamoto,T., Harada,N., Kawano,Y., Taya,S., and Kaibuchi,K. (1999). In vivo interaction of AF-6 with activated Ras and ZO-1. Biochem Biophys Res Commun 259, 103-107. Yamamoto,Y., Nishimura,N., Morimoto,S., Kitamura,H., Manabe,S., Kanayama,H.O., Kagawa,S., and Sasaki,T. (2003). Distinct roles of Rab3B and Rab13 in the polarized transport of apical, basolateral, and tight junctional membrane proteins to the plasma membrane. Biochem Biophys Res Commun 308, 270-275. Yamanaka,T., Horikoshi,Y., Suzuki,A., Sugiyama,Y., Kitamura,K., Maniwa,R., Nagai,Y., Yamashita,A., Hirose,T., Ishikawa,H., and Ohno,S. (2001). PAR-6 regulates aPKC activity in a novel way and mediates cell-cell contact-induced formation of the epithelial junctional complex. Genes Cells 6, 721-731. 222 Yang,Y., Janich,S., Cohn,J.A., and Wilson,J.M. (1993). The common variant of cystic fibrosis transmembrane conductance regulator is recognized by hsp70 and degraded in a pre-Golgi nonlysosomal compartment. Proc Natl Acad Sci U S A 90, 9480-9484. Yap,A.S., Brieher,W.M., Pruschy,M., and Gumbiner,B.M. (1997). Lateral clustering of the adhesive ectodomain: a fundamental determinant of cadherin function. Curr Biol 7, 308-315. Yap,A.S., Niessen,C.M., and Gumbiner,B.M. (1998). The juxtamembrane region of the cadherin cytoplasmic tail supports lateral clustering, adhesive strengthening, and interaction with p120ctn. J Cell Biol 141, 779-789. Yeager,M., Unger,V.M., and Falk,M.M. (1998). Synthesis, assembly and structure of gap junction intercellular channels. Curr Opin Struct. Biol 8, 517-524. Yonemura,S., Itoh,M., Nagafuchi,A., and Tsukita,S. (1995). Cell-to-cell adherens junction formation and actin filament organization: similarities and differences between non-polarized fibroblasts and polarized epithelial cells. J Cell Sci 108 ( Pt 1), 127-142. Young,D., Shprintzen,R.J., and Goldberg,R.B. (1980). Cardiac malformations in the velocardiofacial syndrome. Am J Cardiol. 46, 643-648. Yu,A.L., McCarthy,K.M., Francis,S.A., McCormack,J.M., Lai,J., Rogers,R.A., Lynch,R.D., and Schneeberger,E.E. (2005). Knock Down of Occludin Expression Leads to Diverse Phenotypic Alterations in Epithelial Cells. Am J Physiol Cell Physiol. Zahraoui,A., Joberty,G., Arpin,M., Fontaine,J.J., Hellio,R., Tavitian,A., and Louvard,D. (1994). A small rab GTPase is distributed in cytoplasmic vesicles in non polarized cells but colocalizes with the tight junction marker ZO-1 in polarized epithelial cells. J Cell Biol 124, 101-115. Zahraoui,A., Louvard,D., and Galli,T. (2000). Tight junction, a platform for trafficking and signaling protein complexes. J Cell Biol 151, F31-F36. Zeitlin,P.L., Diener-West,M., Rubenstein,R.C., Boyle,M.P., Lee,C.K., and Brass-Ernst,L. (2002). Evidence of CFTR function in cystic fibrosis after systemic administration of 4phenylbutyrate. Mol Ther. 6, 119-126. Zettl,K.S., Sjaastad,M.D., Riskin,P.M., Parry,G., Machen,T.E., and Firestone,G.L. (1992). Glucocorticoid-induced formation of tight junctions in mouse mammary epithelial cells in vitro. Proc Natl Acad Sci U S A 89, 9069-9073. Zhadanov,A.B., Provance,D.W., Jr., Speer,C.A., Coffin,J.D., Goss,D., Blixt,J.A., Reichert,C.M., and Mercer,J.A. (1999). Absence of the tight junctional protein AF-6 disrupts epithelial cell-cell junctions and cell polarity during mouse development. Curr Biol 9, 880-888. 223 [...]... intermixing of apical and basolateral plasma membrane components TJs consist of integral membrane proteins such as claudin, occludin and JAM proteins that are linked via cytoplasmic peripheral proteins e.g ZO-1, -2 and -3 to the actin cytoskeleton This study identifies the roles of zonula occludens (ZO) proteins and characterizes their interacting partners Novel proteins that interact with ZO-1 were identified... strands represent integral membrane proteins that polymerize linearly within the lipid bilayer with their extracellular domain contacting that of proteins in strands on adjacent cells (Fig 5A) The ‘lipid model’, in contrast, depicts strands as cylindrical micelles with the polar head groups of the lipids directed inward and the hydrophobic tail immersed in the lipid matrix of the plasma membrane of. .. recent identification of TJ integral and peripheral proteins, the ‘protein model’ gained additional support In particular, the identification of the TJ integral proteins, occludin and claudin, and the observation that overexpression of these proteins in MDCK cells increases TER of MDCK cell monolayers (Balda et al., 1996; Van Itallie et al., 2001) strongly implicate a central role for proteins in TJ function... laterally and tightly associates in trans with another TJ-strand in the apposing membrane of an adjacent cell to form a paired strand completely obliterating the intercellular space (Fig 2C) The number of TJ strands as well as the frequency of their ramification vary from one cell type to another and are likely of significance for the functional properties of a particular TJ 18 A B C D Fig 2: Structure of. .. at the level of tight junction 22 1.3: Models of Tight Junctions Two types of models have been proposed to explain the features of TJ strands seen in freeze fractures (Fig 5) TJ strands were proposed to be composed of either proteins (Cereijido et al., 1978; Chalcroft and Bullivant, 1970; Griepp et al., 1983; PolakCharcon et al., 1978) or lipids (Kachar and Reese, 1982; Kan, 1993; Pinto and Kachar,... or regulatory proteins, e.g Rab13, Rab3B, Sec6/8, aPKC, PP2A and PTEN, and (4) Transcriptional and post-transcriptional regulators, e.g symplekin, ZONAB and HuASH Fig 6: The composition of tight junctions Tight junction proteins are composed of integral membrane proteins that are linked to the actin cytoskeleton via peripheral membrane proteins (Figure reprinted from Johnson et al., 2005 with permission... TJ transmembrane proteins Several integral membrane proteins have been identified in recent years and only three classes of integral membrane proteins, namely occludin, claudin and JAM proteins (Fig 7), will be discussed here Claudins and JAMs are protein families, with individual members showing distinct tissue distributions and functions, thus adding to the complexity and uniqueness of tight junctions... an intracellular seal that acts as a barrier and fence to prevent paracellular diffusion and intermixing of proteins and lipids between the apical and basolateral plasma membrane domains, respectively AJs and desmosomes are mechanically linked to AJ and desmosomes on neighboring cells as well as to the actin and intermediate filament cytoskeleton, respectively, providing strength and rigidity to the... model (A) involves integral membrane proteins that polymerize within the lipid bilayers and interacts with proteins on neighboring cells The lipid model (B) involves inverted lipid cylindrical micelles, which constitute TJ strands Figure reproduced with permission from Macmillan Magazines Ltd (Tsukita et al., 2001) 24 1.4: Protein components of TJ The protein components of TJ can be divided in at least... motion of the eyeball Tenany: An abnormal condition characterized by periodic painful muscular spasms and tremors, caused by faulty calcium metabolism and associated with diminished function of the parathyroid glands 13 SUMMARY Zonulae Occludens (ZO) or tight junctions (TJ) are specialized plasma membrane domains that regulate the paracellular transepithelial permeability and prevent the intermixing of . IDENTIFICATION AND CHARACTERIZATION OF PROTEINS THAT INTERACT WITH ZONULA OCCLUDENS PROTEINS P JAYA KAUSALYA INSTITUTE OF MOLECULAR AND CELL BIOLOGY. BIOLOGY NATIONAL UNIVERSITY OF SINGAPORE 2005 2 IDENTIFICATION AND CHARACTERIZATION OF PROTEINS THAT INTERACT WITH ZONULA OCCLUDENS PROTEINS P JAYA KAUSALYA. in TJ that are part of MAGUK family. Fig. 13: Structure of PDZ3 domain of PSD-95 with a peptide ligand. Fig. 14: Possible modes of interaction of PDZ containing proteins. Fig. 15: ZO-3 interacts