Isoform-specific expression of the Coxsackie and adenovirus receptor (CAR) in neuromuscular junction and cardiac intercalated discs

Shaw, Christian A.; Holland, Paul C.; Sinnreich, Michael; Allen, Carol A.; Sollerbrant, Kerstin; Karpati, George; Nalbantoglu, Joséphine

doi:10.1186/1471-2121-5-42

Cited by 46 publications

(26 citation statements)

References 30 publications

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“…The preparation of a His-tagged fusion protein expressing amino acids 259 -339 of the cytoplasmic domain of CAR has been described previously (35). This fusion protein contains the domains that are common to the two full-length CAR isoforms (m1 and m2 (34)) but lacks the distal most 26 amino acids of the m1 isoform, including the PDZ binding residues (SIV).…”

Section: Methodsmentioning

confidence: 99%

The Coxsackie and Adenovirus Receptor Binds Microtubules and Plays a Role in Cell Migration

Fok

Huang

Holland

et al. 2007

Journal of Biological Chemistry

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The Coxsackie and adenovirus receptor (CAR), a cell adhesion molecule of the immunoglobulin superfamily, inhibits cell growth of a variety of tumors. The cytoplasmic domain of CAR has been implicated in decreased invasion and intracerebral growth of human U87 glioma cells. Using affinity binding, we identified tubulin as an interaction partner for the cytoplasmic domain of CAR. The interaction was specific; CAR and tubulin co-immunoprecipitated in cells expressing endogenous CAR and partially co-localized in situ. The binding of CAR to tubulin heterodimers and to microtubules was direct, with dissociation constants of ϳ1 M for tubulin and ϳ32 nM for in vitro assembled microtubules. Whereas CAR-expressing U87 glioma cells had decreased migration in a chemotactic assay in Boyden chambers as compared with control cells, an effect that depended on the presence of the cytoplasmic domain of CAR, the difference was abrogated at low, non-cytotoxic doses of the taxane paclitaxel, a microtubule-stabilizing agent. These results indicate that CAR may affect cell migration through its interaction with microtubules.

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Section: Methodsmentioning

confidence: 99%

The Coxsackie and Adenovirus Receptor Binds Microtubules and Plays a Role in Cell Migration

Fok

Huang

Holland

et al. 2007

Journal of Biological Chemistry

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“…Although differences in localization were primarily observed in liver and kidney tissues, large differences in the relative amount of mCAR-1 protein expression was found between liver, kidney, pancreas and colon, and relative to mCAR-2. Differential isoform localization has also been shown in skeletal muscle, where mCAR-2 is the primary isoform at the neuromuscular junction in mice and both CAR Ex7 and CAR Ex8 are present in human (Shaw et al, 2004). Perhaps the most striking difference in isoform localization has been observed in mature spermatozoa where each isoform was present in a distinct region of the acrosome (Mirza et al, 2006), however, subsequent studies that conditionally knocked out both isoforms in mice did not show any defect in spermatogenesis or fertilization (Sultana et al, 2014).…”

Section: Discussionmentioning

confidence: 95%

Adenovirus transduction: More complicated than receptor expression

2017

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The abundance and accessibility of a primary virus receptor are critical factors that impact the susceptibility of a host cell to virus infection. The Coxsackievirus and adenovirus receptor (CAR) has two transmembrane isoforms that occur due to alternative splicing and differ in localization and function in polarized epithelia. To determine the relevance of isoform-specific expression across cell types, the abundance and localization of both isoforms were determined in ten common cell lines, and correlated with susceptibility to adenovirus transduction relative to polarized primary human airway epithelia. Data show that the gene and protein expression for each isoform of CAR varies significantly between cell lines and polarization, as indicated by high transepithelial resistance, is inversely related to adenovirus transduction. In summary, the variability of polarity and isoform-specific expression among model cells are critical parameters that must be considered when evaluating the clinical relevance of potential adenovirus-mediated gene therapy and anti-adenovirus strategies.

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“…The rabbit polyclonal antibody RP291 raised against the C-terminal intracellular domain of human CAR (46 kDa isoform) cross-reacts with the murine homolog mCAR1 [12,33], and was a kind gift from Dr. Kerstin Sollerbrant (Karolinska Institute and University Hospital, Stockholm, Sweden). Rabbit polyclonal antibody raised against ADAM10 was from AnaSpec, Inc.…”

Section: Methodsmentioning

confidence: 99%

“…As a component of epithelial cell tight junctions, CAR participates in forming a barrier to paracellular flow of macromolecules, binding to tight junction proteins such as zonula occludens-1 (ZO-1) [9] and multi-PDZ domain protein-1 (MUPP-1) [10]. Although CAR expression is high in the developing heart and skeletal muscle, it is barely detectable by adulthood, becoming restricted to skeletal muscle neuromuscular junctions and cardiac intercalated discs [11,12]. CAR expression is critical for normal cardiac development, as its gene deletion in mice before embryonic day 11 results in cardiac abnormalities and embryonic lethality [13–15].…”

Section: Introductionmentioning

confidence: 99%

The Coxsackievirus and Adenovirus Receptor (CAR) Undergoes Ectodomain Shedding and Regulated Intramembrane Proteolysis (RIP)

2013

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The Coxsackievirus and Adenovirus Receptor (CAR) is a cell adhesion molecule originally characterized as a virus receptor but subsequently shown to be involved in physiological processes such as neuronal and heart development, epithelial tight junction integrity, and tumour suppression. Proteolysis of cell adhesion molecules and a wide variety of other cell surface proteins serves as a mechanism for protein turnover and, in some cases, cell signaling. Metalloproteases such as A Disintegrin and Metalloprotease (ADAM) family members cleave cell surface receptors to release their substrates’ ectodomains, while the presenilin/ɣ-secretase complex mediates regulated intramembrane proteolysis (RIP), releasing intracellular domain fragments from the plasma membrane. In the case of some substrates such as Notch and amyloid precursor protein (APP), the released intracellular domains enter the nucleus to modulate gene expression. We report that CAR ectodomain is constitutively shed from glioma cells and developing neurons, and is also shed when cells are treated with the phorbol ester phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore ionomycin. We identified ADAM10 as a sheddase of CAR using assays involving shRNA knockdown and rescue, overexpression of wild-type ADAM10 and inhibition of ADAM10 activity by addition of its prodomain. In vitro peptide cleavage, mass spectrometry and mutagenesis revealed the amino acids M224 to L227 of CAR as the site of ADAM10-mediated ectodomain cleavage. CAR also undergoes RIP by the presenilin/γ-secretase complex, and the intracellular domain of CAR enters the nucleus. Ectodomain shedding is a prerequisite for RIP of CAR. Thus, CAR belongs to the increasing list of cell surface molecules that undergo ectodomain shedding and that are substrates for ɣ-secretase-mediated RIP.

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Isoform-specific expression of the Coxsackie and adenovirus receptor (CAR) in neuromuscular junction and cardiac intercalated discs

Cited by 46 publications

References 30 publications

The Coxsackie and Adenovirus Receptor Binds Microtubules and Plays a Role in Cell Migration

The Coxsackie and Adenovirus Receptor Binds Microtubules and Plays a Role in Cell Migration

Adenovirus transduction: More complicated than receptor expression

The Coxsackievirus and Adenovirus Receptor (CAR) Undergoes Ectodomain Shedding and Regulated Intramembrane Proteolysis (RIP)

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