title: Interspecies differences in virus uptake versus cardiac function of the coxsackievirus and adenovirus receptor.
abstract: The coxsackievirus and adenovirus receptor (CAR) is a cell contact protein with an important role in virus uptake. Its extracellular immunoglobulin domains mediate the binding to coxsackievirus and adenovirus as well as homophilic and heterophilic interactions between cells. The cytoplasmic tail links CAR to the cytoskeleton and intracellular signaling cascades. In the heart, CAR is crucial for embryonic development, electrophysiology, and coxsackievirus B infection. Noncardiac functions are less well understood, in part due to the lack of suitable animal models. Here, we generated a transgenic mouse that rescued the otherwise embryonic-lethal CAR knockout (KO) phenotype by expressing chicken CAR exclusively in the heart. Using this rescue model, we addressed interspecies differences in coxsackievirus uptake and noncardiac functions of CAR. Survival of the noncardiac CAR KO (ncKO) mouse indicates an essential role for CAR in the developing heart but not in other tissues. In adult animals, cardiac activity was normal, suggesting that chicken CAR can replace the physiological functions of mouse CAR in the cardiomyocyte. However, chicken CAR did not mediate virus entry in vivo, so that hearts expressing chicken instead of mouse CAR were protected from infection and myocarditis. Comparison of sequence homology and modeling of the D1 domain indicate differences between mammalian and chicken CAR that relate to the sites important for virus binding but not those involved in homodimerization. Thus, CAR-directed anticoxsackievirus therapy with only minor adverse effects in noncardiac tissue could be further improved by selectively targeting the virus-host interaction while maintaining cardiac function.
  
  
  OBJECTIVE
  Coxsackievirus B3 (CVB3) is one of the most common human pathogens causing myocarditis. Its receptor, the coxsackievirus and adenovirus receptor (CAR), not only mediates virus uptake but also relates to cytoskeletal organization and intracellular signaling. Animals without CAR die prenatally with major cardiac malformations. In the adult heart, CAR is important for virus entry and electrical conduction, but its nonmuscle functions are largely unknown. Here, we show that chicken CAR expression exclusively in the heart can rescue the otherwise embryonic-lethal CAR knockout phenotype but does not support CVB3 infection of adult cardiomyocytes. Our findings have implications for the evolution of virus-host versus physiological interactions involving CAR and could help to improve future coxsackievirus-directed therapies inhibiting virus replication while maintaining CAR's cellular functions.
authors.affiliation: Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Freiberg
authors.affiliation: Department for Molecular Pathology, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Sauter
authors.affiliation: Department of Applied Biochemistry, University of Technology, Berlin, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Pinkert
authors.affiliation: Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Thirupugal
authors.initials: T
authors.lastname: Govindarajan
authors.affiliation: Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Joanna
authors.initials: J
authors.lastname: Kaldrack
authors.affiliation: Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Meghna
authors.initials: M
authors.lastname: Thakkar
authors.affiliation: Department of Applied Biochemistry, University of Technology, Berlin, Germany.
authors.forename: Henry
authors.initials: H
authors.lastname: Fechner
authors.affiliation: Department for Molecular Pathology, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Klingel
authors.affiliation: Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, Germany gotthardt@mdc-berlin.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Gotthardt
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002645:Chickens; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein / Q000502:physiology*; D003384:Coxsackievirus Infections / Q000517:prevention & control* / Q000821:virology; D029822:Enterovirus B, Human / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006321:Heart / Q000502:physiology* / Q000821:virology; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009205:Myocarditis / Q000517:prevention & control* / Q000821:virology; D014779:Virus Replication
chemical_list: D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein
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