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

This Article Full Text Full Text (PDF, 1225K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Buchert, M. Articles by Hovens, C. M. Search for Related Content PubMed PubMed Citation Articles by Buchert, M. Articles by Hovens, C. M. Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/1999//361 $5.00
The Journal of Cell Biology, Volume 144, Number 2, , 1999 361-371

The Junction-associated Protein AF-6 Interacts and Clusters with Specific Eph Receptor Tyrosine Kinases at Specialized Sites of Cell–Cell Contact in the Brain

Michael Buchert^*, Stefan Schneider^*, Virginia Meskenaite, Mark T. Adams^*, Eli Canaani^¶, Thomas Baechi, Karin Moelling^*, and Christopher M. Hovens^*

^* Institut für Medizinische Virologie and Elektronenmikroskopisches Zentrallabor, Universität Zürich; Institut für Neuroinformatik, Universität Zürich/Eidgenössische Technische Hochschule, CH-8028 Zürich, Switzerland; and ^¶ Weizmann Institute for Science, Rehovot 76100, Israel

The AF-6/afadin protein, which contains a single PDZ domain, forms a peripheral component of cell membranes at specialized sites of cell–cell junctions. To identify potential receptor-binding targets of AF-6 we screened the PDZ domain of AF-6 against a range of COOH-terminal peptides selected from receptors having potential PDZ domain-binding termini. The PDZ domain of AF-6 interacts with a subset of members of the Eph subfamily of RTKs via its COOH terminus both in vitro and in vivo. Cotransfection of a green fluorescent protein-tagged AF-6 fusion protein with full-length Eph receptors into heterologous cells induces a clustering of the Eph receptors and AF-6 at sites of cell–cell contact. Immunohistochemical analysis in the adult rat brain reveals coclustering of AF-6 with Eph receptors at postsynaptic membrane sites of excitatory synapses in the hippocampus. Furthermore, AF-6 is a substrate for a subgroup of Eph receptors and phosphorylation of AF-6 is dependent on a functional kinase domain of the receptor. The physical interaction of endogenous AF-6 with Eph receptors is demonstrated by coimmunoprecipitation from whole rat brain lysates. AF-6 is a candidate for mediating the clustering of Eph receptors at postsynaptic specializations in the adult rat brain.

Key Words: postsynaptic clustering • PDZ domains • receptor tyrosine kinases • neuron physiology • Ras-binding protein

Abbreviations used in this paper: aa, amino acid; FLAG AF-6 PDZ, FLAG epitope-tagged AF-6 PDZ domain construct; GFP, green fluorescent protein; GST, glutathione-S-transferase; MAGUK, membrane-associated guanylate kinase; NGS, normal goat serum; NMDA, N-methyl-D-aspartate; NR2, NMDA receptor 2; PB, phosphate buffer; RIPA, radioimmunoprecipitation assay; RT, room temperature; RTK, receptor tyrosine kinase; SAP, synapse-associated protein.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• Giagtzoglou, N., Ly, C. V., Bellen, H. J. (2009). Cell Adhesion, the Backbone of the Synapse: "Vertebrate" and "Invertebrate" Perspectives. Cold Spring Harb. Perspect. Biol. 1: a003079-a003079 [Abstract] [Full Text]  
• North, H. A., Zhao, X., Kolk, S. M., Clifford, M. A., Ziskind, D. M., Donoghue, M. J. (2009). Promotion of proliferation in the developing cerebral cortex by EphA4 forward signaling. Development 136: 2467-2476 [Abstract] [Full Text]  
• Xie, Z., Photowala, H., Cahill, M. E., Srivastava, D. P., Woolfrey, K. M., Shum, C. Y., Huganir, R. L., Penzes, P. (2008). Coordination of Synaptic Adhesion with Dendritic Spine Remodeling by AF-6 and Kalirin-7. J. Neurosci. 28: 6079-6091 [Abstract] [Full Text]  
• Richter, M., Murai, K. K., Bourgin, C., Pak, D. T., Pasquale, E. B. (2007). The EphA4 Receptor Regulates Neuronal Morphology through SPAR-Mediated Inactivation of Rap GTPases. J. Neurosci. 27: 14205-14215 [Abstract] [Full Text]  
• Lorger, M., Moelling, K. (2006). Regulation of epithelial wound closure and intercellular adhesion by interaction of AF6 with actin cytoskeleton. J. Cell Sci. 119: 3385-3398 [Abstract] [Full Text]  
• Zhang, Z., Rehmann, H., Price, L. S., Riedl, J., Bos, J. L. (2005). AF6 Negatively Regulates Rap1-induced Cell Adhesion. J. Biol. Chem. 280: 33200-33205 [Abstract] [Full Text]  
• Matsuoka, H., Obama, H., Kelly, M. L., Matsui, T., Nakamoto, M. (2005). Biphasic Functions of the Kinase-defective Ephb6 Receptor in Cell Adhesion and Migration. J. Biol. Chem. 280: 29355-29363 [Abstract] [Full Text]  
• Ivanov, A. I., Steiner, A. A., Scheck, A. C., Romanovsky, A. A. (2005). Expression of Eph receptors and their ligands, ephrins, during lipopolysaccharide fever in rats. Physiol. Genomics 21: 152-160 [Abstract] [Full Text]  
• Zhou, H., Xu, Y., Yang, Y., Huang, A., Wu, J., Shi, Y. (2005). Solution Structure of AF-6 PDZ Domain and Its Interaction with the C-terminal Peptides from Neurexin and Bcr. J. Biol. Chem. 280: 13841-13847 [Abstract] [Full Text]  
• Calo, L., Bruno, V., Spinsanti, P., Molinari, G., Korkhov, V., Esposito, Z., Patane, M., Melchiorri, D., Freissmuth, M., Nicoletti, F. (2005). Interactions between Ephrin-B and Metabotropic Glutamate 1 Receptors in Brain Tissue and Cultured Neurons. J. Neurosci. 25: 2245-2254 [Abstract] [Full Text]  
• Sato, T., Kanai, Y., Noma, T., Kanai-Azuma, M., Taya, S., Matsui, T., Ishii, M., Kawakami, H., Kurohmaru, M., Kaibuchi, K., Wood, S. A, Hayashi, Y. (2004). A close correlation in the expression patterns of Af-6 and Usp9x in Sertoli and granulosa cells of mouse testis and ovary. Reproduction 128: 583-594 [Abstract] [Full Text]  
• Murai, K. K., Pasquale, E. B. (2004). Eph Receptors, Ephrins, and Synaptic Function. Neuroscientist 10: 304-314 [Abstract]  
• Lee, N. P.Y., Cheng, C. Y. (2004). Adaptors, Junction Dynamics, and Spermatogenesis. Biol. Reprod. 71: 392-404 [Abstract] [Full Text]  
• Henkemeyer, M., Itkis, O. S., Ngo, M., Hickmott, P. W., Ethell, I. M. (2003). Multiple EphB receptor tyrosine kinases shape dendritic spines in the hippocampus. JCB 163: 1313-1326 [Abstract] [Full Text]  
• Boettner, B., Harjes, P., Ishimaru, S., Heke, M., Fan, H. Q., Qin, Y., Van Aelst, L., Gaul, U. (2003). The AF-6 Homolog Canoe Acts as a Rap1 Effector During Dorsal Closure of the Drosophila Embryo. Genetics 165: 159-169 [Abstract] [Full Text]  
• Bundesen, L. Q., Scheel, T. A., Bregman, B. S., Kromer, L. F. (2003). Ephrin-B2 and EphB2 Regulation of Astrocyte-Meningeal Fibroblast Interactions in Response to Spinal Cord Lesions in Adult Rats. J. Neurosci. 23: 7789-7800 [Abstract] [Full Text]  
• Radziwill, G., Erdmann, R. A., Margelisch, U., Moelling, K. (2003). The Bcr Kinase Downregulates Ras Signaling by Phosphorylating AF-6 and Binding to Its PDZ Domain. Mol. Cell. Biol. 23: 4663-4672 [Abstract] [Full Text]  
• Nourry, C., Grant, S. G. N., Borg, J.-P. (2003). PDZ Domain Proteins: Plug and Play!. Sci Signal 2003: re7-re7 [Abstract] [Full Text]  
• Takai, Y., Nakanishi, H. (2003). Nectin and afadin: novel organizers of intercellular junctions. J. Cell Sci. 116: 17-27 [Abstract] [Full Text]  
• Prevost, N., Woulfe, D., Tanaka, T., Brass, L. F. (2002). Interactions between Eph kinases and ephrins provide a mechanism to support platelet aggregation once cell-to-cell contact has occurred. Proc. Natl. Acad. Sci. USA 99: 9219-9224 [Abstract] [Full Text]  
• Trivier, E., Ganesan, T. S. (2002). RYK, a Catalytically Inactive Receptor Tyrosine Kinase, Associates with EphB2 and EphB3 but Does Not Interact with AF-6. J. Biol. Chem. 277: 23037-23043 [Abstract] [Full Text]  
• Mizoguchi, A., Nakanishi, H., Kimura, K., Matsubara, K., Ozaki-Kuroda, K., Katata, T., Honda, T., Kiyohara, Y., Heo, K., Higashi, M., Tsutsumi, T., Sonoda, S., Ide, C., Takai, Y. (2002). Nectin: an adhesion molecule involved in formation of synapses. JCB 156: 555-565 [Abstract] [Full Text]  
• Reymond, N., Fabre, S., Lecocq, E., Adelaide, J., Dubreuil, P., Lopez, M. (2001). Nectin4/PRR4, a New Afadin-associated Member of the Nectin Family That Trans-interacts with Nectin1/PRR1 through V Domain Interaction. J. Biol. Chem. 276: 43205-43215 [Abstract] [Full Text]  
• Vaccaro, P., Brannetti, B., Montecchi-Palazzi, L., Philipp, S., Citterich, M. H., Cesareni, G., Dente, L. (2001). Distinct Binding Specificity of the Multiple PDZ Domains of INADL, a Human Protein with Homology to INAD from Drosophila melanogaster. J. Biol. Chem. 276: 42122-42130 [Abstract] [Full Text]  
• Tao, H. W., Poo, M.-m. (2001). Retrograde signaling at central synapses. Proc. Natl. Acad. Sci. USA 98: 11009-11015 [Abstract] [Full Text]  
• Zhang, W., Benson, D. L. (2001). Stages of Synapse Development Defined by Dependence on F-Actin. J. Neurosci. 21: 5169-5181 [Abstract] [Full Text]  
• Yi, H., Leunissen, J. L.M., Shi, G.-M., Gutekunst, C.-A., Hersch, S. M. (2001). A Novel Procedure for Pre-embedding Double Immunogold-Silver Labeling at the Ultrastructural Level. J. Histochem. Cytochem. 49: 279-284 [Abstract] [Full Text]  
• Kuwada, N., Kimura, F., Matsumura, T., Yamashita, T., Nakamura, Y., Wakimoto, N., Ikeda, T., Sato, K., Motoyoshi, K. (2001). t(11;14)(q23;q24) Generates an MLL-Human Gephyrin Fusion Gene along with a de facto Truncated MLL in Acute Monoblastic Leukemia. Cancer Res. 61: 2665-2669 [Abstract] [Full Text]  
• Maudsley, S., Zamah, A. M., Rahman, N., Blitzer, J. T., Luttrell, L. M., Lefkowitz, R. J., Hall, R. A. (2000). Platelet-Derived Growth Factor Receptor Association with Na+/H+ Exchanger Regulatory Factor Potentiates Receptor Activity. Mol. Cell. Biol. 20: 8352-8363 [Abstract] [Full Text]  
• Tachibana, K., Nakanishi, H., Mandai, K., Ozaki, K., Ikeda, W., Yamamoto, Y., Nagafuchi, A., Tsukita, S., Takai, Y. (2000). Two Cell Adhesion Molecules, Nectin and Cadherin, Interact through Their Cytoplasmic Domain-Associated Proteins. JCB 150: 1161-1176 [Abstract] [Full Text]  
• Satoh-Horikawa, K., Nakanishi, H., Takahashi, K., Miyahara, M., Nishimura, M., Tachibana, K., Mizoguchi, A., Takai, Y. (2000). Nectin-3, a New Member of Immunoglobulin-like Cell Adhesion Molecules That Shows Homophilic and Heterophilic Cell-Cell Adhesion Activities. J. Biol. Chem. 275: 10291-10299 [Abstract] [Full Text]  
• Thanos, C. D., Faham, S., Goodwill, K. E., Cascio, D., Phillips, M., Bowie, J. U. (1999). Monomeric Structure of the Human EphB2 Sterile alpha Motif Domain. J. Biol. Chem. 274: 37301-37306 [Abstract] [Full Text]  
• Ikeda, W., Nakanishi, H., Miyoshi, J., Mandai, K., Ishizaki, H., Tanaka, M., Togawa, A., Takahashi, K., Nishioka, H., Yoshida, H., Mizoguchi, A., Nishikawa, S.-i., Takai, Y. (1999). Afadin: A Key Molecule Essential for Structural Organization of Cell-Cell Junctions of Polarized Epithelia during Embryogenesis. JCB 146: 1117-1132 [Abstract] [Full Text]  
• Gale, N. W., Yancopoulos, G. D. (1999). Growth factors acting via endothelial cell-specific receptor tyrosine kinases: VEGFs, Angiopoietins, and ephrins in vascular development. Genes Dev. 13: 1055-1066 [Full Text]  
• Takasu, M. A., Dalva, M. B., Zigmond, R. E., Greenberg, M. E. (2002). Modulation of NMDA Receptor- Dependent Calcium Influx and Gene Expression Through EphB Receptors. Science 295: 491-495 [Abstract] [Full Text]  
• Ebnet, K., Schulz, C. U., Meyer zu Brickwedde, M.-K., Pendl, G. G., Vestweber, D. (2000). Junctional Adhesion Molecule Interacts with the PDZ Domain-containing Proteins AF-6 and ZO-1. J. Biol. Chem. 275: 27979-27988 [Abstract] [Full Text]  
• Ligensa, T., Krauss, S., Demuth, D., Schumacher, R., Camonis, J., Jaques, G., Weidner, K. M. (2001). A PDZ Domain Protein Interacts with the C-terminal Tail of the Insulin-like Growth Factor-1 Receptor but Not with the Insulin Receptor. J. Biol. Chem. 276: 33419-33427 [Abstract] [Full Text]  
• Mizoguchi, A., Nakanishi, H., Kimura, K., Matsubara, K., Ozaki-Kuroda, K., Katata, T., Honda, T., Kiyohara, Y., Heo, K., Higashi, M., Tsutsumi, T., Sonoda, S., Ide, C., Takai, Y. (2002). Nectin: an adhesion molecule involved in formation of synapses. JCB 156: 555-565 [Abstract] [Full Text]  

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