Infection of cultured mammalian cells with African horse sickness virus (AHSV) is known to induce cell death. To date, the trigger(s) of this response, the apoptotic pathways activated during AHSV infection and the functional consequences of apoptosis on the virus replication cycle have yet to be characterized. This study demonstrated that extracellular treatment of BHK-21 cells with both of the AHSV4 outer capsid proteins, VP2 and VP5, was sufficient to trigger apoptosis. Whether steps in AHSV4 replication subsequent to viral attachment were required for AHSV4-induced apoptosis was also investigated. Apoptosis was induced in BHK-21 cells infected with UV-inactivated AHSV4 and in ribavirin-treated cells infected with AHSV4. However, both AHSV4- and VP2/VP5-stimulated apoptotic responses were inhibited in the presence of the endosomal acidification inhibitors ammonium chloride and chloroquine. These results indicated that uncoating of AHSV4 virions, but not viral transcription or subsequent steps in viral replication, was required for AHSV4 to induce apoptosis in BHK-21 cells. Furthermore, this study showed that both the extrinsic (caspase-8) and intrinsic (caspase-9) apoptotic pathways were induced following AHSV4 infection. The inhibition of caspase activity in AHSV4-infected cells did not diminish AHSV4 replication, but reduced the release and dissemination of progeny viral particles. Taken together, the data indicated that uncoating of AHSV virions was required for apoptosis induction, and that apoptosis enhanced virus spread and release.
AhmadM.,
SrinivasulaS.M.,
WangL.,
TalanianR.V.,
LitwackG.,
Fernandes-AlnemriT.,
AlnemriE.S.
(1997). CRADD, a novel human apoptotic adaptor molecule for caspase-2, and FasL/tumor necrosis factor receptor-interacting protein RIPCancer Res57615–619.
[Google Scholar]
AshkenaziA.,
DixitV.M.
(1998). Death receptors: signaling and modulationScience2811305–1308.[CrossRef][Google Scholar]
BestS.M.
(2008). Viral subversion of apoptotic enzymes: escape from death rowAnnu Rev Microbiol62171–192.[CrossRef][Google Scholar]
CarrascosaA.L.,
BustosM.J.,
NogalM.L.,
González de BuitragoG.,
RevillaY.
(2002). Apoptosis induced in an early step of African swine fever virus entry into vero cells does not require virus replicationVirology294372–382.[CrossRef][Google Scholar]
ClarkeP.,
DebiasiR.L.,
GoodyR.,
HoytC.C.,
Richardson-BurnsS.,
TylerK.L.
(2005). Mechanisms of reovirus-induced cell death and tissue injury: role of apoptosis and virus-induced perturbation of host-cell signaling and transcription factor activationViral Immunol1889–115.[CrossRef][Google Scholar]
CoetzerJ.A.W.,
GuthrieA.J.
(2004). African horse sickness. In Infectious Diseases of Livestock, pp. 1231–1246. Edited by
CoetzerJ. A. W.,
TustinR. C.
Cape TownOxford University Press.
[Google Scholar]
ConnollyJ.L.,
DermodyT.S.
(2002). Virion disassembly is required for apoptosis induced by reovirusJ Virol761632–1641.[CrossRef][Google Scholar]
DanthiP.
(2011). Enter the kill zone: initiation of death signaling during virus entryVirology411316–324.[CrossRef][Google Scholar]
DanthiP.,
HansbergerM.W.,
CampbellJ.A.,
ForrestJ.C.,
DermodyT.S.
(2006). JAM-A-independent, antibody-mediated uptake of reovirus into cells leads to apoptosisJ Virol801261–1270.[CrossRef][Google Scholar]
DanthiP.,
CoffeyC.M.,
ParkerJ.S.,
AbelT.W.,
DermodyT.S.
(2008). Independent regulation of reovirus membrane penetration and apoptosis by the μ1 φ domainPLoS Pathog4e1000248.[CrossRef][Google Scholar]
DeMaulaC.D.,
JutilaM.A.,
WilsonD.W.,
MacLachlanN.J.
(2001). Infection kinetics, prostacyclin release and cytokine-mediated modulation of the mechanism of cell death during bluetongue virus infection of cultured ovine and bovine pulmonary artery and lung microvascular endothelial cellsJ Gen Virol82787–794.
[Google Scholar]
DispersynG.,
NuydensR.,
ConnorsR.,
BorgersM.,
GeertsH.
(1999). Bcl-2 protects against FCCP-induced apoptosis and mitochondrial membrane potential depolarization in PC12 cellsBiochim Biophys Acta1428357–371.[CrossRef][Google Scholar]
DuC.,
FangM.,
LiY.,
LiL.,
WangX.
(2000). Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibitionCell10233–42.[CrossRef][Google Scholar]
GalluzziL.,
KeppO.,
MorselliE.,
VitaleI.,
SenovillaL.,
PintiM.,
ZitvogelL.,
KroemerG.
(2010). Viral strategies for the evasion of immunogenic cell deathJ Intern Med267526–542.[CrossRef][Google Scholar]
Gómez-VillamandosJ.C.,
SánchezC.,
CarrascoL.,
LaviadaM.M.,
BautistaM.J.,
Martínez-TorrecuadradaJ.,
Sánchez-VizcaínoJ.M.,
SierraM.A.
(1999). Pathogenesis of African horse sickness: ultrastructural study of the capillaries in experimental infectionJ Comp Pathol121101–116.[CrossRef][Google Scholar]
GraciJ.D.,
CameronC.E.
(2006). Mechanisms of action of ribavirin against distinct virusesRev Med Virol1637–48.[CrossRef][Google Scholar]
HassanS.S.,
RoyP.
(1999). Expression and functional characterization of bluetongue virus VP2 protein: role in cell entryJ Virol739832–9842.
[Google Scholar]
HassanS.H.,
WirblichC.,
ForzanM.,
RoyP.
(2001). Expression and functional characterization of bluetongue virus VP5 protein: role in cellular permeabilizationJ Virol758356–8367.[CrossRef][Google Scholar]
HayS.,
KannourakisG.
(2002). A time to kill: viral manipulation of the cell death programJ Gen Virol831547–1564.
[Google Scholar]
IsakovicA.,
JankovicT.,
HarhajiL.,
Kostic-RajacicS.,
NikolicZ.,
VajsV.,
TrajkovicV.
(2008). Antiglioma action of xanthones from Gentiana kochiana: mechanistic and structure-activity requirementsBioorg Med Chem165683–5694.[CrossRef][Google Scholar]
JanJ.T.,
GriffinD.E.
(1999). Induction of apoptosis by Sindbis virus occurs at cell entry and does not require virus replicationJ Virol7310296–10302.
[Google Scholar]
KerrJ.F.R.,
WyllieA.H.,
CurrieA.R.
(1972). Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kineticsBr J Cancer26239–257.[CrossRef][Google Scholar]
KominskyD.J.,
BickelR.J.,
TylerK.L.
(2002). Reovirus-induced apoptosis requires both death receptor- and mitochondrial-mediated caspase-dependent pathways of cell deathCell Death Differ9926–933.[CrossRef][Google Scholar]
KoyamaA.H.,
AdachiA.
(1997). Induction of apoptosis by herpes simplex virus type 1J Gen Virol782909–2912.
[Google Scholar]
LabradaL.,
BodelónG.,
ViñuelaJ.,
BenaventeJ.
(2002). Avian reoviruses cause apoptosis in cultured cells: viral uncoating, but not viral gene expression, is required for apoptosis inductionJ Virol767932–7941.[CrossRef][Google Scholar]
LaegreidW.W.,
BurrageT.G.,
Stone-MarschatM.,
SkowronekA.
(1992). Electron microscopic evidence for endothelial infection by African horsesickness virusVet Pathol29554–556.[CrossRef][Google Scholar]
LecatsasG.
(1968). Electron microscopic study of the formation of bluetongue virusOnderstepoort J Vet Res35139–149.
[Google Scholar]
LecatsasG.,
ErasmusB.J.
(1967). Electron microscopic study of the formation of African horse-sickness virusArch Gesamte Virusforsch22442–450.[CrossRef][Google Scholar]
MarkovicZ.,
Todorovic-MarkovicB.,
KleutD.,
NikolicN.,
Vranjes-DjuricS.,
MisirkicM.,
VucicevicL.,
JanjetovicK.,
IsakovicA.,
other authors.
(2007). The mechanism of cell-damaging reactive oxygen generation by colloidal fullerenesBiomaterials285437–5448.[CrossRef][Google Scholar]
Martínez-TorrecuadradaJ.L.,
IwataH.,
VenteoA.,
CasalI.,
RoyP.
(1994). Expression and characterization of the two outer capsid proteins of African horsesickness virus: the role of VP2 in virus neutralizationVirology202348–359.[CrossRef][Google Scholar]
MatsuyamaS.,
LlopisJ.,
DeverauxQ.L.,
TsienR.Y.,
ReedJ.C.
(2000). Changes in intramitochondrial and cytosolic pH: early events that modulate caspase activation during apoptosisNat Cell Biol2318–325.[CrossRef][Google Scholar]
MortolaE.,
NoadR.,
RoyP.
(2004). Bluetongue virus outer capsid proteins are sufficient to trigger apoptosis in mammalian cellsJ Virol782875–2883.[CrossRef][Google Scholar]
O'BrienV.
(1998). Viruses and apoptosisJ Gen Virol791833–1845.
[Google Scholar]
O'DonnellS.M.,
HansbergerM.W.,
ConnollyJ.L.,
ChappellJ.D.,
WatsonM.J.,
PierceJ.M.,
WetzelJ.D.,
HanW.,
BartonE.S.,
other authors.
(2005). Organ-specific roles for transcription factor NF-kappaB in reovirus-induced apoptosis and diseaseJ Clin Invest1152341–2350.[CrossRef][Google Scholar]
OellermannR.A.
(1970). Plaque formation by African horsesickness virus and characterization of its RNAOnderstepoort J Vet Res37137–143.
[Google Scholar]
OsawaY.,
HazratiA.
(1965). Growth of African horse sickness virus in monkey kidney cell culturesAm J Vet Res25505–511.
[Google Scholar]
RankinJ.T.Jr,
EppesS.B.,
AntczakJ.B.,
JoklikW.K.
(1989). Studies on the mechanism of the antiviral activity of ribavirin against reovirusVirology168147–158.[CrossRef][Google Scholar]
Rodríguez-GrilleJ.,
BuschL.K.,
Martínez-CostasJ.,
BenaventeJ.
(2014). Avian reovirus-triggered apoptosis enhances both virus spread and the processing of the viral nonstructural muNS proteinVirology462-46349–59.[CrossRef][Google Scholar]
RoulstonA.,
MarcellusR.C.,
BrantonP.E.
(1999). Viruses and apoptosisAnnu Rev Microbiol53577–628.[CrossRef][Google Scholar]
SambrookJ.,
RussellD.W.
(2001). Molecular Cloning: A Laboratory Manual3rd edn.Cold Spring Harbour, NYCold Spring Harbour Laboratory.
[Google Scholar]
ShaiB.,
SchmuklerE.,
YanivR.,
ZivN.,
HornG.,
BumbarovV.,
YadinH.,
SmorodinskyN.I.,
BacharachE.,
other authors.
(2013). Epizootic hemorrhagic disease virus induces and benefits from cell stress, autophagy, and apoptosisJ Virol8713397–13408.[CrossRef][Google Scholar]
StassenL.,
HuismansH.,
TheronJ.
(2011). Membrane permeabilization of the African horse sickness virus VP5 protein is mediated by two N-terminal amphipathic α-helicesArch Virol156711–715.[CrossRef][Google Scholar]
StewartM.E.,
RoyP.
(2010). Role of cellular caspases, nuclear factor-kappa B and interferon regulatory factors in Bluetongue virus infection and cell fateVirol J7362.[CrossRef][Google Scholar]
SusinS.A.,
LorenzoH.K.,
ZamzamiN.,
MarzoI.,
SnowB.E.,
BrothersG.M.,
MangionJ.,
JacototE.,
CostantiniP.,
other authors.
(1999). Molecular characterization of mitochondrial apoptosis-inducing factorNature397441–446.[CrossRef][Google Scholar]
TurkB.,
TurkD.,
TurkV.
(2000). Lysosomal cysteine proteases: more than scavengersBiochim Biophys Acta147798–111.[CrossRef][Google Scholar]
VenterE.
(2014). Factors contributing to AHSV pathogenesis in infected mammalian and insect cellsPhD thesis, University of Pretoria, Pretoria, South Africa.
[Google Scholar]
ZhangX.,
BoyceM.,
BhattacharyaB.,
ZhangX.,
ScheinS.,
RoyP.,
ZhouZ.H.
(2010). Bluetongue virus coat protein VP2 contains sialic acid-binding domains, and VP5 resembles enveloped virus fusion proteinsProc Natl Acad Sci U S A1076292–6297.[CrossRef][Google Scholar]
ZouH.,
LiY.,
LiuX.,
WangX.
(1999). An APAF-1.cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9J Biol Chem27411549–11556.[CrossRef][Google Scholar]