title: Mutations in VP1 and 3A proteins improve binding and replication of rhinovirus C15 in HeLa-E8 cells.
abstract: Viruses in the rhinovirus C species (RV-C) can cause severe respiratory illnesses in children including pneumonia and asthma exacerbations. A transduced cell line (HeLa-E8) stably expressing the CDHR3-Y529receptor variant, supports propagation of RV-C after infection. C15 clinical or recombinant isolates replicate in HeLa-E8, however progeny yields are lower than those of related strains of RV-A and RV-B. Serial passaging of C15 in HeLa-E8 resulted in stronger cytopathic effects and increased (≥10-fold) virus binding to cells and progeny yields. The adaptation was acquired by two mutations which increased binding (VP1 T125K) and replication (3A E41K), respectively. A similar 3A mutation engineered into C2 and C41 cDNAs also improved viral replication (2-8 fold) in HeLa but the heparan sulfate mediated cell-binding enhancement by the VP1 change was C15-specific. The findings now enable large-scale cost-effective C15 production by infection and the testing of RV-C infectivity by plaque assay.
authors.affiliation: Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA. Electronic address: yabochkov@wisc.edu.
authors.forename: Yury A
authors.initials: YA
authors.lastname: Bochkov
authors.affiliation: Institute for Molecular Virology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Watters
authors.affiliation: Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Sarmila
authors.initials: S
authors.lastname: Basnet
authors.affiliation: Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Shakher
authors.initials: S
authors.lastname: Sijapati
authors.affiliation: Institute for Molecular Virology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Marchel
authors.initials: M
authors.lastname: Hill
authors.affiliation: Institute for Molecular Virology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Ann C
authors.initials: AC
authors.lastname: Palmenberg
authors.affiliation: Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA; Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Gern
mesh_terms: D020816:Amino Acid Motifs; D006367:HeLa Cells; D006801:Humans; D020125:Mutation, Missense; D010850:Picornaviridae Infections / Q000821:virology*; D012229:Rhinovirus / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D053585:Virus Attachment; D014779:Virus Replication
chemical_list: D014764:Viral Proteins; C074995:viral protein 1, rhinovirus
keywords: Adaptation; Heparan sulfate; Pathogenesis; Receptor specificity; Rhinovirus C