title: Single amino acid changes in the virus capsid permit coxsackievirus B3 to bind decay-accelerating factor.
abstract: Many coxsackievirus B isolates bind to human decay-accelerating factor (DAF) as well as to the coxsackievirus and adenovirus receptor (CAR). The first-described DAF-binding isolate, coxsackievirus B3 (CB3)-RD, was obtained during passage of the prototype strain CB3-Nancy on RD cells, which express DAF but very little CAR. CB3-RD binds to human DAF, whereas CB3-Nancy does not. To determine the molecular basis for the specific interaction of CB3-RD with DAF, we produced cDNA clones encoding both CB3-RD and CB3-Nancy and mutated each of the sites at which the RD and Nancy sequences diverged. We found that a single amino acid change, the replacement of a glutamate within VP3 (VP3-234E) with a glutamine residue (Q), conferred upon CB3-Nancy the capacity to bind DAF and to infect RD cells. Readaptation of molecularly cloned CB3-Nancy to RD cells selected for a new virus with the same VP3-234Q residue. In experiments with CB3-H3, another virus isolate that does not bind measurably to DAF, adaptation to RD cells resulted in a DAF-binding isolate with a single amino acid change within VP2 (VP2-138 N to D). Both VP3-234Q and VP2-138D were required for binding of CB3-RD to DAF. In the structure of the CB3-RD-DAF complex determined by cryo-electron microscopy, both VP3-234Q and VP2-138D are located at the contact site between the virus and DAF.
authors.affiliation: The Children's Hospital of Philadelphia, 3615 Civic Center Blvd., Philadelphia, PA 19104, USA.
authors.forename: Jieyan
authors.initials: J
authors.lastname: Pan
authors.forename: Bhargavi
authors.initials: B
authors.lastname: Narayanan
authors.forename: Shardule
authors.initials: S
authors.lastname: Shah
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Yoder
authors.forename: Javier O
authors.initials: JO
authors.lastname: Cifuente
authors.forename: Susan
authors.initials: S
authors.lastname: Hafenstein
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Bergelson
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001483:Base Sequence; D002213:Capsid; D017931:DNA Primers; D029822:Enterovirus B, Human / Q000737:chemistry / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers