Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus

Adam Taylor, Julian Melton, Lara Herrero, Bastian Thaa, Liis Karo-Astover, Peter Gage, Michelle Nelson, Kuo-Ching Sheng, Brett Lidbury, Gary Ewart, Gerald McInerney, Andres Merits, Suresh MAHALINGAM

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The alphaviral 6k gene 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 the 6k proteins in the arthritogenic alphavirus Ross River virus (RRV) in infected cells and in mice, using a novel 6k in-frame deletion mutant. Comprehensive microscopic analysis revealed that the 6k proteins were predominantly localized at the endoplasmic reticulum of RRV-infected cells. RRV virions that lack the 6k proteins 6K and TF [RRV-(¿6K)] were more vulnerable to changes in pH, and the corresponding virus had increased sensitivity to a higher temperature. While the 6k deletion did not reduce RRV particle production in BHK-21 cells, it affected virion release from the host cell. Subsequent in vivo studies demonstrated that RRV-(¿6K) caused a milder disease than wild-type virus, with viral titers being reduced in infected mice. Immunization of mice with RRV-(¿6K) 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 the 6k proteins may contribute to alphaviral disease manifestations and suggest that manipulation of the 6k gene may be a potential strategy to facilitate viral vaccine development.
Original languageEnglish
Pages (from-to)4150-4159
Number of pages10
JournalJournal of Virology
Volume90
Issue number8
DOIs
Publication statusPublished - 2016

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Ross River virus
gene deletion
Gene Deletion
Genome
loci
genome
Proteins
virion
Virion
Alphavirus
proteins
viral load
mice
Ribosomal Frameshifting
Chikungunya virus
Viral Vaccines
cells
viruses
structural proteins
vaccine development

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Taylor, A., Melton, J., Herrero, L., Thaa, B., Karo-Astover, L., Gage, P., ... MAHALINGAM, S. (2016). Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus. Journal of Virology, 90(8), 4150-4159. https://doi.org/10.1128/JVI.03192-15
Taylor, Adam ; Melton, Julian ; Herrero, Lara ; Thaa, Bastian ; Karo-Astover, Liis ; Gage, Peter ; Nelson, Michelle ; Sheng, Kuo-Ching ; Lidbury, Brett ; Ewart, Gary ; McInerney, Gerald ; Merits, Andres ; MAHALINGAM, Suresh. / Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus. In: Journal of Virology. 2016 ; Vol. 90, No. 8. pp. 4150-4159.
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abstract = "The alphaviral 6k gene 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 the 6k proteins in the arthritogenic alphavirus Ross River virus (RRV) in infected cells and in mice, using a novel 6k in-frame deletion mutant. Comprehensive microscopic analysis revealed that the 6k proteins were predominantly localized at the endoplasmic reticulum of RRV-infected cells. RRV virions that lack the 6k proteins 6K and TF [RRV-(¿6K)] were more vulnerable to changes in pH, and the corresponding virus had increased sensitivity to a higher temperature. While the 6k deletion did not reduce RRV particle production in BHK-21 cells, it affected virion release from the host cell. Subsequent in vivo studies demonstrated that RRV-(¿6K) caused a milder disease than wild-type virus, with viral titers being reduced in infected mice. Immunization of mice with RRV-(¿6K) 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 the 6k proteins may contribute to alphaviral disease manifestations and suggest that manipulation of the 6k gene may be a potential strategy to facilitate viral vaccine development.",
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Taylor, A, Melton, J, Herrero, L, Thaa, B, Karo-Astover, L, Gage, P, Nelson, M, Sheng, K-C, Lidbury, B, Ewart, G, McInerney, G, Merits, A & MAHALINGAM, S 2016, 'Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus', Journal of Virology, vol. 90, no. 8, pp. 4150-4159. https://doi.org/10.1128/JVI.03192-15

Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus. / Taylor, Adam; Melton, Julian; Herrero, Lara; Thaa, Bastian; Karo-Astover, Liis; Gage, Peter; Nelson, Michelle; Sheng, Kuo-Ching; Lidbury, Brett; Ewart, Gary; McInerney, Gerald; Merits, Andres; MAHALINGAM, Suresh.

In: Journal of Virology, Vol. 90, No. 8, 2016, p. 4150-4159.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of an in-frame deletion of the 6k gene locus from the genome of Ross River virus

AU - Taylor, Adam

AU - Melton, Julian

AU - Herrero, Lara

AU - Thaa, Bastian

AU - Karo-Astover, Liis

AU - Gage, Peter

AU - Nelson, Michelle

AU - Sheng, Kuo-Ching

AU - Lidbury, Brett

AU - Ewart, Gary

AU - McInerney, Gerald

AU - Merits, Andres

AU - MAHALINGAM, Suresh

N1 - Copyright © 2016, American Society for Microbiology. All Rights Reserved.

PY - 2016

Y1 - 2016

N2 - The alphaviral 6k gene 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 the 6k proteins in the arthritogenic alphavirus Ross River virus (RRV) in infected cells and in mice, using a novel 6k in-frame deletion mutant. Comprehensive microscopic analysis revealed that the 6k proteins were predominantly localized at the endoplasmic reticulum of RRV-infected cells. RRV virions that lack the 6k proteins 6K and TF [RRV-(¿6K)] were more vulnerable to changes in pH, and the corresponding virus had increased sensitivity to a higher temperature. While the 6k deletion did not reduce RRV particle production in BHK-21 cells, it affected virion release from the host cell. Subsequent in vivo studies demonstrated that RRV-(¿6K) caused a milder disease than wild-type virus, with viral titers being reduced in infected mice. Immunization of mice with RRV-(¿6K) 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 the 6k proteins may contribute to alphaviral disease manifestations and suggest that manipulation of the 6k gene may be a potential strategy to facilitate viral vaccine development.

AB - The alphaviral 6k gene 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 the 6k proteins in the arthritogenic alphavirus Ross River virus (RRV) in infected cells and in mice, using a novel 6k in-frame deletion mutant. Comprehensive microscopic analysis revealed that the 6k proteins were predominantly localized at the endoplasmic reticulum of RRV-infected cells. RRV virions that lack the 6k proteins 6K and TF [RRV-(¿6K)] were more vulnerable to changes in pH, and the corresponding virus had increased sensitivity to a higher temperature. While the 6k deletion did not reduce RRV particle production in BHK-21 cells, it affected virion release from the host cell. Subsequent in vivo studies demonstrated that RRV-(¿6K) caused a milder disease than wild-type virus, with viral titers being reduced in infected mice. Immunization of mice with RRV-(¿6K) 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 the 6k proteins may contribute to alphaviral disease manifestations and suggest that manipulation of the 6k gene may be a potential strategy to facilitate viral vaccine development.

UR - http://www.scopus.com/inward/record.url?scp=84963864965&partnerID=8YFLogxK

U2 - 10.1128/JVI.03192-15

DO - 10.1128/JVI.03192-15

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SP - 4150

EP - 4159

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

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ER -