The role of IKKβ in Venezuelan equine encephalitis virus infection.

Venezuelan equine encephalitis virus (VEEV) belongs to the genus Alphavirus, family Togaviridae. VEEV infection is characterized by extensive inflammation and studies from other laboratories implicated an involvement of the NF-κB cascade in the in vivo pathology. Initial studies indicated that at ea...

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Main Authors: Moushimi Amaya, Kelsey Voss, Gavin Sampey, Svetlana Senina, Cynthia de la Fuente, Claudius Mueller, Valerie Calvert, Kylene Kehn-Hall, Calvin Carpenter, Fatah Kashanchi, Charles Bailey, Soren Mogelsvang, Emanuel Petricoin, Aarthi Narayanan
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0086745&type=printable
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author Moushimi Amaya
Kelsey Voss
Gavin Sampey
Svetlana Senina
Cynthia de la Fuente
Claudius Mueller
Valerie Calvert
Kylene Kehn-Hall
Calvin Carpenter
Fatah Kashanchi
Charles Bailey
Soren Mogelsvang
Emanuel Petricoin
Aarthi Narayanan
author_facet Moushimi Amaya
Kelsey Voss
Gavin Sampey
Svetlana Senina
Cynthia de la Fuente
Claudius Mueller
Valerie Calvert
Kylene Kehn-Hall
Calvin Carpenter
Fatah Kashanchi
Charles Bailey
Soren Mogelsvang
Emanuel Petricoin
Aarthi Narayanan
author_sort Moushimi Amaya
collection DOAJ
description Venezuelan equine encephalitis virus (VEEV) belongs to the genus Alphavirus, family Togaviridae. VEEV infection is characterized by extensive inflammation and studies from other laboratories implicated an involvement of the NF-κB cascade in the in vivo pathology. Initial studies indicated that at early time points of VEEV infection, the NF-κB complex was activated in cells infected with the TC-83 strain of VEEV. One upstream kinase that contributes to the phosphorylation of p65 is the IKKβ component of the IKK complex. Our previous studies with Rift valley fever virus, which exhibited early activation of the NF-κB cascade in infected cells, had indicated that the IKKβ component underwent macromolecular reorganization to form a novel low molecular weight form unique to infected cells. This prompted us to investigate if the IKK complex undergoes a comparable macromolecular reorganization in VEEV infection. Size-fractionated VEEV infected cell extracts indicated a macromolecular reorganization of IKKβ in VEEV infected cells that resulted in formation of lower molecular weight complexes. Well-documented inhibitors of IKKβ function, BAY-11-7082, BAY-11-7085 and IKK2 compound IV, were employed to determine whether IKKβ function was required for the production of infectious progeny virus. A decrease in infectious viral particles and viral RNA copies was observed with inhibitor treatment in the attenuated and virulent strains of VEEV infection. In order to further validate the requirement of IKKβ for VEEV replication, we over-expressed IKKβ in cells and observed an increase in viral titers. In contrast, studies carried out using IKKβ(-/-) cells demonstrated a decrease in VEEV replication. In vivo studies demonstrated that inhibitor treatment of TC-83 infected mice increased their survival. Finally, proteomics studies have revealed that IKKβ may interact with the viral protein nsP3. In conclusion, our studies have revealed that the host IKKβ protein may be critically involved in VEEV replication.
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spelling doaj-art-cd6a2ad0145f41758f9ef5a709426b2c2025-08-20T03:11:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8674510.1371/journal.pone.0086745The role of IKKβ in Venezuelan equine encephalitis virus infection.Moushimi AmayaKelsey VossGavin SampeySvetlana SeninaCynthia de la FuenteClaudius MuellerValerie CalvertKylene Kehn-HallCalvin CarpenterFatah KashanchiCharles BaileySoren MogelsvangEmanuel PetricoinAarthi NarayananVenezuelan equine encephalitis virus (VEEV) belongs to the genus Alphavirus, family Togaviridae. VEEV infection is characterized by extensive inflammation and studies from other laboratories implicated an involvement of the NF-κB cascade in the in vivo pathology. Initial studies indicated that at early time points of VEEV infection, the NF-κB complex was activated in cells infected with the TC-83 strain of VEEV. One upstream kinase that contributes to the phosphorylation of p65 is the IKKβ component of the IKK complex. Our previous studies with Rift valley fever virus, which exhibited early activation of the NF-κB cascade in infected cells, had indicated that the IKKβ component underwent macromolecular reorganization to form a novel low molecular weight form unique to infected cells. This prompted us to investigate if the IKK complex undergoes a comparable macromolecular reorganization in VEEV infection. Size-fractionated VEEV infected cell extracts indicated a macromolecular reorganization of IKKβ in VEEV infected cells that resulted in formation of lower molecular weight complexes. Well-documented inhibitors of IKKβ function, BAY-11-7082, BAY-11-7085 and IKK2 compound IV, were employed to determine whether IKKβ function was required for the production of infectious progeny virus. A decrease in infectious viral particles and viral RNA copies was observed with inhibitor treatment in the attenuated and virulent strains of VEEV infection. In order to further validate the requirement of IKKβ for VEEV replication, we over-expressed IKKβ in cells and observed an increase in viral titers. In contrast, studies carried out using IKKβ(-/-) cells demonstrated a decrease in VEEV replication. In vivo studies demonstrated that inhibitor treatment of TC-83 infected mice increased their survival. Finally, proteomics studies have revealed that IKKβ may interact with the viral protein nsP3. In conclusion, our studies have revealed that the host IKKβ protein may be critically involved in VEEV replication.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0086745&type=printable
spellingShingle Moushimi Amaya
Kelsey Voss
Gavin Sampey
Svetlana Senina
Cynthia de la Fuente
Claudius Mueller
Valerie Calvert
Kylene Kehn-Hall
Calvin Carpenter
Fatah Kashanchi
Charles Bailey
Soren Mogelsvang
Emanuel Petricoin
Aarthi Narayanan
The role of IKKβ in Venezuelan equine encephalitis virus infection.
PLoS ONE
title The role of IKKβ in Venezuelan equine encephalitis virus infection.
title_full The role of IKKβ in Venezuelan equine encephalitis virus infection.
title_fullStr The role of IKKβ in Venezuelan equine encephalitis virus infection.
title_full_unstemmed The role of IKKβ in Venezuelan equine encephalitis virus infection.
title_short The role of IKKβ in Venezuelan equine encephalitis virus infection.
title_sort role of ikkβ in venezuelan equine encephalitis virus infection
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0086745&type=printable
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