title: Internally deleted WNV genomes isolated from exotic birds in New Mexico: function in cells, mosquitoes, and mice.
abstract: Most RNA viruses exist in their hosts as a heterogeneous population of related variants. Due to error prone replication, mutants are constantly generated which may differ in individual fitness from the population as a whole. Here we characterize three WNV isolates that contain, along with full-length genomes, mutants with large internal deletions to structural and nonstructural protein-coding regions. The isolates were all obtained from lorikeets that died from WNV at the Rio Grande Zoo in Albuquerque, NM between 2005 and 2007. The deletions are approximately 2kb, in frame, and result in the elimination of the complete envelope, and portions of the prM and NS-1 proteins. In Vero cell culture, these internally deleted WNV genomes function as defective interfering particles, reducing the production of full-length virus when introduced at high multiplicities of infection. In mosquitoes, the shortened WNV genomes reduced infection and dissemination rates, and virus titers overall, and were not detected in legs or salivary secretions at 14 or 21 days post-infection. In mice, inoculation with internally deleted genomes did not attenuate pathogenesis relative to full-length or infectious clone derived virus, and shortened genomes were not detected in mice at the time of death. These observations provide evidence that large deletions may occur within flavivirus populations more frequently than has generally been appreciated and suggest that they impact population phenotype minimally. Additionally, our findings suggest that highly similar mutants may frequently occur in particular vertebrate hosts.
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, NM, USA. Kendra.Pesko@yale.edu
authors.forename: Kendra N
authors.initials: KN
authors.lastname: Pesko
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Fitzpatrick
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Ryan
authors.forename: Pei-Yong
authors.initials: PY
authors.lastname: Shi
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Niall J
authors.initials: NJ
authors.lastname: Lennon
authors.forename: Ruchi M
authors.initials: RM
authors.lastname: Newman
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Henn
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Ebel
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001717:Birds / Q000821:virology; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D009033:Culicidae / Q000821:virology; D003673:Defective Viruses / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D016679:Genome, Viral; D007668:Kidney / Q000166:cytology / Q000378:metabolism / Q000821:virology; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009154:Mutation / Q000235:genetics; D009516:New Mexico; D012367:RNA, Viral / Q000302:isolation & purification; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics; D014902:West Nile virus / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D012367:RNA, Viral; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; C074932:prM protein, Flavivirus