The PDZ3 domain of the cellular scaffolding protein MAGI-1 interacts with the Coxsackievirus and adenovirus receptor (CAR) |
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| Institution: | 1. Department of Biological Sciences, Wright State University, Dayton, OH 45435, USA;2. Department of Biochemistry and Molecular Biology, Wright State University, Dayton, OH 45435, USA;1. Dokuz Eylul University, Faculty of Medicine, Department of Medical Biology, Inciralti, 35340 Izmir, Turkey;2. Dokuz Eylul University, Izmir Biomedicine and Genome Center, Inciralti, 35340 Izmir, Turkey;3. Dokuz Eylul University, Faculty of Medicine, Department of Pharmacology, Inciralti, 35340 Izmir, Turkey;1. British Heart Foundation Centre for Cardiovascular Science, College of Medicine and Veterinary Medicine, University of Edinburgh, Room Suite 305, The Chancellor''s Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK;2. Imaging Science Laboratories, Department of Radiology, Translational and Molecular Imaging Institute, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029, USA;3. Imaging Science Laboratories, Department of Medicine, Translational and Molecular Imaging Institute, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029, USA;1. Department of Pediatrics, University of Florida, Gainesville, Florida, USA;2. Powell Gene Therapy Center, University of Florida, Gainesville, Florida, USA;3. Department of Physical Therapy, University of Florida, Gainesville, Florida, USA |
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| Abstract: | The Coxsackievirus and adenovirus receptor (CAR) is an essential cellular protein that is involved in cell–cell adhesion, protein trafficking, and viral infection. The major isoform of CAR is selectively sorted to the basolateral membrane of polarized epithelial cells where it co-localizes with the cellular scaffolding protein membrane-associated guanylate kinase with inverted domain structure-1 (MAGI-1). Previously, we demonstrated CAR interacts with MAGI-1 through a PDZ–domain dependent interaction. Here, we show that the PDZ3 domain of MAGI-1 is exclusively responsible for the high affinity interaction between the seven exon isoform of CAR and MAGI-1 using yeast-two-hybrid analysis and confirming this interaction biochemically and in cellular lysates by in vitro pull down assay and co-immunoprecipitation. The high affinity interaction between the PDZ3 domain and CAR C-terminus was measured by fluorescence resonance energy transfer. Further, we investigated the biological relevance of this high affinity interaction between CAR and the PDZ3 domain of MAGI-1 and found that it does not alter CAR-mediated adenovirus infection. By contrast, interruption of this high affinity interaction altered the localization of MAGI-1 indicating that CAR is able to traffic MAGI-1 to cell junctions. These data deepen the molecular understanding of the interaction between CAR and MAGI-1 and indicate that although CAR plays a role in trafficking PDZ-based scaffolding proteins to cellular junctions, association with a high affinity intracellular binding partner does not significantly alter adenovirus binding and entry via CAR. |
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| Keywords: | Adenovirus Epithelia Cell junction PDZ domain |
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