title: A single-amino-acid polymorphism in Chikungunya virus E2 glycoprotein influences glycosaminoglycan utilization.
abstract: Chikungunya virus (CHIKV) is a reemerging arbovirus responsible for outbreaks of infection throughout Asia and Africa, causing an acute illness characterized by fever, rash, and polyarthralgia. Although CHIKV infects a broad range of host cells, little is known about how CHIKV binds and gains access to the target cell interior. In this study, we tested whether glycosaminoglycan (GAG) binding is required for efficient CHIKV replication using CHIKV vaccine strain 181/25 and clinical isolate SL15649. Preincubation of strain 181/25, but not SL15649, with soluble GAGs resulted in dose-dependent inhibition of infection. While parental Chinese hamster ovary (CHO) cells are permissive for both strains, neither strain efficiently bound to or infected mutant CHO cells devoid of GAG expression. Although GAGs appear to be required for efficient binding of both strains, they exhibit differential requirements for GAGs, as SL15649 readily infected cells that express excess chondroitin sulfate but that are devoid of heparan sulfate, whereas 181/25 did not. We generated a panel of 181/25 and SL15649 variants containing reciprocal amino acid substitutions at positions 82 and 318 in the E2 glycoprotein. Reciprocal exchange at residue 82 resulted in a phenotype switch; Gly(82) results in efficient infection of mutant CHO cells but a decrease in heparin binding, whereas Arg(82) results in reduced infectivity of mutant cells and an increase in heparin binding. These results suggest that E2 residue 82 is a primary determinant of GAG utilization, which likely mediates attenuation of vaccine strain 181/25.
  
  
  OBJECTIVE
  Chikungunya virus (CHIKV) infection causes a debilitating rheumatic disease that can persist for months to years, and yet there are no licensed vaccines or antiviral therapies. Like other alphaviruses, CHIKV displays broad tissue tropism, which is thought to be influenced by virus-receptor interactions. In this study, we determined that cell-surface glycosaminoglycans are utilized by both a vaccine strain and a clinical isolate of CHIKV to mediate virus binding. We also identified an amino acid polymorphism in the viral E2 attachment protein that influences utilization of glycosaminoglycans. These data enhance an understanding of the viral and host determinants of CHIKV cell entry, which may foster development of new antivirals that act by blocking this key step in viral infection.
authors.affiliation: Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Silva
authors.forename: Solomiia
authors.initials: S
authors.lastname: Khomandiak
authors.forename: Alison W
authors.initials: AW
authors.lastname: Ashbrook
authors.forename: Romy
authors.initials: R
authors.lastname: Weller
authors.forename: Mark T
authors.initials: MT
authors.lastname: Heise
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Morrison
authors.forename: Terence S
authors.initials: TS
authors.lastname: Dermody
mesh_terms: D018354:Alphavirus Infections / Q000378:metabolism; D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D065632:Chikungunya Fever; D002646:Chikungunya virus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D011992:Endosomes / Q000378:metabolism / Q000821:virology; D005838:Genotype; D006025:Glycosaminoglycans / Q000378:metabolism* / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D055503:Protein Multimerization; D055672:Static Electricity; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics*; D053585:Virus Attachment / Q000187:drug effects; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D014759:Viral Envelope Proteins