title: Effects of an In-Frame Deletion of the 6k Gene Locus from the Genome of Ross River Virus.
abstract: The alphaviral6kgene region encodes the two structural proteins 6K protein and, due to a ribosomal frameshift event, the transframe protein (TF). Here, we characterized the role of the6kproteins in the arthritogenic alphavirus Ross River virus (RRV) in infected cells and in mice, using a novel6kin-frame deletion mutant. Comprehensive microscopic analysis revealed that the6kproteins were predominantly localized at the endoplasmic reticulum of RRV-infected cells. RRV virions that lack the6kproteins 6K and TF [RRV-(Δ6K)] were more vulnerable to changes in pH, and the corresponding virus had increased sensitivity to a higher temperature. While the6kdeletion did not reduce RRV particle production in BHK-21 cells, it affected virion release from the host cell. Subsequentin vivostudies demonstrated that RRV-(Δ6K) caused a milder disease than wild-type virus, with viral titers being reduced in infected mice. Immunization of mice with RRV-(Δ6K) resulted in a reduced viral load and accelerated viral elimination upon secondary infection with wild-type RRV or another alphavirus, chikungunya virus (CHIKV). Our results show that the6kproteins may contribute to alphaviral disease manifestations and suggest that manipulation of the6kgene may be a potential strategy to facilitate viral vaccine development.
  
  
  OBJECTIVE
  Arthritogenic alphaviruses, such as chikungunya virus (CHIKV) and Ross River virus (RRV), cause epidemics of debilitating rheumatic disease in areas where they are endemic and can emerge in new regions worldwide. RRV is of considerable medical significance in Australia, where it is the leading cause of arboviral disease. The mechanisms by which alphaviruses persist and cause disease in the host are ill defined. This paper describes the phenotypic properties of an RRV6kdeletion mutant. The absence of the6kgene reduced virion release from infected cells and also reduced the severity of disease and viral titers in infected mice. Immunization with the mutant virus protected mice against viremia not only upon exposure to RRV but also upon challenge with CHIKV. These findings could lead to the development of safer and more immunogenic alphavirus vectors for vaccine delivery.
authors: Emerging Viruses and Inflammation Research Group, Institute for Glycomics, Griffith University, Gold Coast, QLD, Australia.
authors: Adam
authors: A
authors: Taylor
authors: Division of Molecular Biosciences, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
authors: Julian V
authors: JV
authors: Melton
authors: Emerging Viruses and Inflammation Research Group, Institute for Glycomics, Griffith University, Gold Coast, QLD, Australia.|Virus and Inflammation Research Group, Faculty of Applied Science, University of Canberra, Canberra, ACT, Australia.
authors: Lara J
authors: http://orcid.org/0000-0002-5661-9828
authors: LJ
authors: Herrero
authors: Karolinska Institutet, Department of Microbiology, Tumor, and Cell Biology (MTC), Stockholm, Sweden.
authors: Bastian
authors: B
authors: Thaa
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Liis
authors: L
authors: Karo-Astover
authors: Division of Molecular Biosciences, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
authors: Peter W
authors: PW
authors: Gage
authors: Virus and Inflammation Research Group, Faculty of Applied Science, University of Canberra, Canberra, ACT, Australia.
authors: Michelle A
authors: MA
authors: Nelson
authors: Emerging Viruses and Inflammation Research Group, Institute for Glycomics, Griffith University, Gold Coast, QLD, Australia.
authors: Kuo-Ching
authors: KC
authors: Sheng
authors: Department of Genome Biology, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
authors: Brett A
authors: BA
authors: Lidbury
authors: Division of Molecular Biosciences, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
authors: Gary D
authors: GD
authors: Ewart
authors: Karolinska Institutet, Department of Microbiology, Tumor, and Cell Biology (MTC), Stockholm, Sweden.
authors: Gerald M
authors: GM
authors: McInerney
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Andres
authors: A
authors: Merits
authors: Emerging Viruses and Inflammation Research Group, Institute for Glycomics, Griffith University, Gold Coast, QLD, Australia s.mahalingam@griffith.edu.au.|Virus and Inflammation Research Group, Faculty of Applied Science, University of Canberra, Canberra, ACT, Australia.
authors: Suresh
authors: http://orcid.org/0000-0003-3141-8410
authors: S
authors: Mahalingam
mesh_terms: D018354:Alphavirus Infections / Q000276:immunology / Q000503:physiopathology / Q000821:virology*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002646:Chikungunya virus / Q000276:immunology; D002522:Chlorocebus aethiops; D006224:Cricetinae; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D009154:Mutation; D016364:Reading Frames; D012398:Ross River virus / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D017384:Sequence Deletion; D014709:Vero Cells; D019562:Viral Load; D015678:Viral Structural Proteins / Q000032:analysis / Q000235:genetics*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014779:Virus Replication
chemical_list: D015678:Viral Structural Proteins; D014765:Viral Vaccines