Functional Effects of Coxsackievirus and Adenovirus Receptor Glycosylation on Homophilic Adhesion and Adenoviral Infection

Excoffon, Katherine J. D. A.; Gansemer, Nicholas D.; Traver, Geri; Zabner, Joseph

doi:10.1128/jvi.02562-06

Cited by 34 publications

(30 citation statements)

References 31 publications

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“…In contrast, glycosylation appears to inhibit PV binding to PVR (16,26). CAR has two glycosylation sites, one on each extracellular domain (38). He et al (7) observed that a CAR-D1 domain, produced in bacteria and therefore lacking glycosylation, bound poorly to CVB3 (7), and other authors (38) have suggested that this might indicate a role for glycosylation.…”

Section: Discussionmentioning

confidence: 99%

“…CAR has two glycosylation sites, one on each extracellular domain (38). He et al (7) observed that a CAR-D1 domain, produced in bacteria and therefore lacking glycosylation, bound poorly to CVB3 (7), and other authors (38) have suggested that this might indicate a role for glycosylation. However, we found that deglycosylation of the D1 and/or D2 domains of CAR had no adverse effect on CVB3 binding or infec- , and an intracellular domain (ICD).…”

Section: Discussionmentioning

confidence: 99%

“…Earlier work demonstrated that the CAR transmembrane and cytoplasmic domains are not required for CVB3 infection (7,27), but the role of D2 had not been investigated. Further, although extracellular glycosylation is known to be important for other CAR-mediated functions, including homotypic adhesion and adenovirus infection (38), the role of glycosylation in CVB3 infection had not been examined. We found here that glycosylation of the extracellular portion of CAR does not contribute to CVB3 infection.…”

Section: Discussionmentioning

confidence: 99%

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The Coxsackievirus and Adenovirus Receptor: Glycosylation and the Extracellular D2 Domain Are Not Required for Coxsackievirus B3 Infection

Pinkert

Röger

Kurreck

et al. 2016

J Virol

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The coxsackievirus and adenovirus receptor (CAR) is a member of the immunoglobulin superfamily (IgSF) and functions as a receptor for coxsackie B viruses (CVBs). The extracellular portion of CAR comprises two glycosylated immunoglobulin-like domains, D1 and D2. CAR-D1 binds to the virus and is essential for virus infection; however, it is not known whether D2 is also important for infection, and the role of glycosylation has not been explored. To understand the function of these structural components in CAR-mediated CVB3 infection, we generated a panel of human (h) CAR deletion and substitution mutants and analyzed their functionality as CVB receptors, examining both virus binding and replication. Lack of glycosylation of the CAR-D1 or -D2 domains did not adversely affect CVB3 binding or infection, indicating that the glycosylation of CAR is not required for its receptor functions. Deletion of the D2 domain reduced CVB3 binding, with a proportionate reduction in the efficiency of virus infection. Replacement of D2 with the homologous D2 domain from chicken CAR, or with the heterologous type C2 immunoglobulin-like domain from IgSF11, another IgSF member, fully restored receptor function; however, replacement of CAR-D2 with domains from CD155 or CD80 restored function only in part. These data indicate that glycosylation of the extracellular domain of hCAR plays no role in CVB3 receptor function and that CAR-D2 is not specifically required. The D2 domain may function largely as a spacer permitting virus access to D1; however, the data may also suggest that D2 affects virus binding by influencing the conformation of D1. IMPORTANCE An important step in virus infection C oxsackie B viruses (CVBs) initiate infection of their host cells by interaction with the coxsackievirus and adenovirus receptor (CAR). An additional cell surface protein, decay-accelerating factor (DAF), promotes binding to the cell surface but is not sufficient for infection (1-3). CAR is a member of the immunoglobulin superfamily (IgSF) and is composed of two extracellular immunoglobulin-like domains, D1 (amino acid [aa] 20 to 139) and D2 (aa 142 to 229), as well as a typical hydrophobic transmembrane domain (TMD; aa 236 to 258) and an internal cytoplasmic domain (ICD; aa 259 to 365) (4). The extracellular immunoglobulin-like domains vary in their secondary structure by different ␤-strand folding. Whereas D1 shows a typical V-type fold structure, D2 has a C2-type immunoglobulin fold (5, 6). Several studies indicate a primary role for the D1 domain in CAR interactions with CVB3 (7) and adenovirus (8), as well as in CAR/CAR homophilic interactions (9-11). Although the isolated D1 domain, produced in Escherichia coli, binds adenovirus efficiently (8), the same D1 domain was found to bind poorly to CVB3 (7), suggesting a possible supporting role for the D2 domain during CAR/ CVB3 interaction.Numerous picornavirus receptors are members of the IgSF: intercellular adhesion molecule-1 (ICAM-1) is a receptor for coxsackievirus A21 (CAV21) and the ma...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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The Coxsackievirus and Adenovirus Receptor: Glycosylation and the Extracellular D2 Domain Are Not Required for Coxsackievirus B3 Infection

Pinkert

Röger

Kurreck

et al. 2016

J Virol

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“…Horseradish peroxidase-labeled secondary antibodies were from Jackson ImmunoResearch (West Grove, PA). Rabbit anti-CAR-1605 and anti-CAR Ex8 -5678 have previously been described (10,11,14,28 (42). For Fluorescence Resonance Energy Transfer (FRET) and binding assays, MAGI-1 PDZ1, PDZ2, and PDZ3 domains and the CAR Ex8 C terminus were cloned (In-Fusion; Clontech) into a pGEX-6p vector (GE Healthcare, Piscataway, NJ) modified to contain a His tag upstream of the glutathione transferase (GST) tag (pHH2).…”

Section: Methodsmentioning

confidence: 99%

The PDZ1 and PDZ3 Domains of MAGI-1 Regulate the Eight-Exon Isoform of the Coxsackievirus and Adenovirus Receptor

Kolawole¹,

Sharma²,

Yan³

et al. 2012

J Virol

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cEpithelial integrity is essential for homeostasis and poses a formidable barrier to pathogen entry. Major factors for viral entry into epithelial cells are the localization and abundance of the primary receptor. The coxsackievirus and adenovirus receptor (CAR) is a primary receptor for these two pathogenic groups of viruses. In polarized epithelia, a low-abundance, alternatively spliced eight-exon isoform of CAR, CAR Ex8 , is localized apically where it can support viral infection from the air-exposed surface. Using biochemical, cell biology, genetic, and spectroscopic approaches, we show that the levels of apical CAR Ex8 are negatively regulated by the PDZ domain-containing protein MAGI-1 (membrane-associated guanylate kinase with inverted orientation protein-1) and that two MAGI-1 PDZ domains, PDZ1 and PDZ3, regulate CAR Ex8 levels in opposing ways. Similar to fulllength MAGI-1, expression of the isolated PDZ3 domain significantly reduces cell surface CAR Ex8 abundance and adenovirus infection. In contrast, the PDZ1 domain is able to rescue CAR Ex8 and adenovirus infection from MAGI-1-mediated suppression. These data suggest a novel cell-based strategy to either suppress viral infection or augment adenovirus-based gene therapy.

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“…The JAM family of proteins has been categorized as belonging to the tight junction, but we classify these proteins as part of the adherens proteins because they are located inferior to the tight junctions (4). We have found that opening the tight junctions with EDTA greatly enhances adenovirus and reovirus interactions with CAR (13). Anchoring proteins, such as cingulin, link JAM proteins to the actin cytoskeleton via their PDZ-binding domain.…”

Section: Adherens Junctionsmentioning

confidence: 99%

Do Cell Junction Protein Mutations Cause an Airway Phenotype in Mice or Humans?

Chang¹,

Pezzulo²,

Zabner³

2011

Am J Respir Cell Mol Biol

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Cell junction proteins connect epithelial cells to each other and to the basement membrane. Genetic mutations of these proteins can cause alterations in some epithelia leading to varied phenotypes such as deafness, renal disease, skin disorders, and cancer. This review examines if genetic mutations in these proteins affect the function of lung airway epithelia. We review cell junction proteins with examples of disease mutation phenotypes in humans and in mouse knockout models. We also review which of these genes are expressed in airway epithelium by microarray expression profiling and immunocytochemistry. Last, we present a comprehensive literature review to find the lung phenotype when cell junction and adhesion genes are mutated or subject to targeted deletion. We found that in murine models, targeted deletion of cell junction and adhesion genes rarely result in a lung phenotype. Moreover, mutations in these genes in humans have no obvious lung phenotype. Our research suggests that simply because a cell junction or adhesion protein is expressed in an organ does not imply that it will exhibit a drastic phenotype when mutated. One explanation is that because a functioning lung is critical to survival, redundancy in the system is expected. Therefore mutations in a single gene might be compensated by a related function of a similar gene product. Further studies in human and animal models will help us understand the overlap in the function of cell junction gene products. Finally, it is possible that the human lung phenotype is subtle and has not yet been described.

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Functional Effects of Coxsackievirus and Adenovirus Receptor Glycosylation on Homophilic Adhesion and Adenoviral Infection

Cited by 34 publications

References 31 publications

The Coxsackievirus and Adenovirus Receptor: Glycosylation and the Extracellular D2 Domain Are Not Required for Coxsackievirus B3 Infection

The Coxsackievirus and Adenovirus Receptor: Glycosylation and the Extracellular D2 Domain Are Not Required for Coxsackievirus B3 Infection

The PDZ1 and PDZ3 Domains of MAGI-1 Regulate the Eight-Exon Isoform of the Coxsackievirus and Adenovirus Receptor

Do Cell Junction Protein Mutations Cause an Airway Phenotype in Mice or Humans?

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