2FNC1†Present address: MRC Protein Phosphorylation Unit, Division of Cell Signalling, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
Feline calicivirus (FCV) is responsible for an acute upper respiratory tract disease in cats. The FCV capsid protein is synthesized as a precursor (76 kDa) that is post-translationally processed into the mature 62 kDa capsid protein by removal of the N-terminal 124 amino acids. Our previous studies have also detected a 40 kDa protein, related to the FCV capsid protein, produced during infection. Here we demonstrate that cleavage of the FCV capsid protein, during infection of cells in culture, was prevented by caspase inhibitors. In addition, caspase-2, -3 and -7 were activated during FCV infection, as shown by pro-form processing, an increase in N-acetyl-Asp-Glu-Val-Asp-7-amido-4-trifluoromethylcoumarin cleavage activity and in situ poly(ADP-ribose) polymerase cleavage. Caspase activation coincided with the induction of apoptosis and capsid cleavage to the 40 kDa fragment. An in vitro cleavage assay, using recombinant human caspases and in vitro-derived FCV capsid protein, revealed that caspase-2, and to a lesser extent caspase-6, cleaved the capsid protein to generate a 40 kDa fragment. Taken together, these results suggest that FCV triggers apoptosis within infected cells and that caspase-induced capsid cleavage occurs concomitantly with apoptosis. The possible role of capsid cleavage in the pathogenesis of FCV infection is discussed.
AlonsoC.,
OviedoJ. M.,
Martin-AlonsoJ. M.,
DiazE.,
BogaJ. A.,
ParraF.1998; Programmed cell death in the pathogenesis of rabbit hemorrhagic disease. Arch Virol 143:321–332
ButtA. J.,
HarveyN. L.,
ParasivamG.,
KumarS.1998; Dimerization and autoprocessing of the Nedd2 (caspase-2) precursor requires both the prodomain and carboxyl-terminal regions. J Biol Chem 273:6763–6768
CarterM. J.,
MiltonI. D.,
MeangerJ.,
BennettM.,
GaskellR. M.,
TurnerP. C.1992a; The complete nucleotide sequence of a feline calicivirus. Virology 190:443–448
CarterM. J.,
MiltonI. D.,
TurnerP. C.,
MeangerJ.,
BennettM.,
GaskellR. M.1992b; Identification and sequence determination of the capsid protein gene of feline calicivirus. Arch Virol 122:223–235
DuvergerV.,
SartoriusU.,
Kelin-BauernschmittP.,
KrammerP. H.,
SchlehoferJ. R.2002; Enhancement of cisplatin-induced apoptosis by infection with adeno-associated virus type 2. Int J Cancer 97:706–712
EleouetJ.-F.,
SleeE. A.,
SauriniF.,
CastagneN.,
PoncetD.,
GarridoC.,
SolaryE.,
MartinS. J.2000; The viral nucleocapsid protein of transmissible gastroenteritis coronavirus (TGEV) is cleaved by caspase-6 and -7 during TGEV-induced apoptosis. J Virol 74:3975–3983
EnariM.,
SakahiraH.,
YokoyamaH.,
OkawaK.,
IwamatsuA.,
NagataS.1998; A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50
GervaisF. G.,
ThornberryN. A.,
RuffoloS. C.,
NicholsonD. W.,
RoyS.1998; Caspases cleave focal adhesion kinase during apoptosis to generate a FRNK-like polypeptide. J Biol Chem 273:17102–17108
HeB.,
LinG. Y.,
DurbinJ. E.,
DurbinR. K.,
LambR. A.2001; The SH integral membrane protein of the paramyxovirus simian virus 5 is required to block apoptosis in MDBK cells. J Virol 75:4068–4079
HerbertT. P.,
BrierleyI.,
BrownT. D. K.1996; Detection of the ORF3 polypeptide of feline calicivirus in infected cells and evidence for its expression from a single, functionally bicistronic, subgenomic mRNA. J Gen Virol 77:123–127
HirataH.,
TakahashiA.,
KobayashiS.,
YoneharaS.,
SawaiH.,
OkazakiT.,
YamamotoK.,
SasadaM.1998; Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis. J Exp Med 187:587–600
JonesR. A.,
JohnsonV. L.,
HintonR. H.,
PoirierG. G.,
ChowS. C.,
KassG. E. N.1999; Liver poly(ADP-ribose)polymerase is resistant to cleavage by caspases. Biochem Biophys Res Commun 256:436–441
KaufmannS. H.1989; Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs – a cautionary note. Cancer Res 49:5870–5878
KumarS.,
KinoshitaM.,
NodaM.,
CopelandN. G.,
JenkinsN. A.1994; Induction of apoptosis by the mouse NEDD2 gene, which encodes a protein similar to the product of the Caenorhabditis elegans cell-death gene CED-3 and the mammalian IL-1- β -converting enzyme. Genes Dev 8:1613–1626
LassusP.,
Opitz-ArayaX.,
LazebnikY.2002; Requirement for caspase-2 in stress-induced apoptosis before mitochondrial permeabilisation. Science 297:1352–1354
LazebnikY. A.,
KaufmannS. H.,
DesnoyersS.,
PoirierG. G.,
EarnshawW. C.1994; Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371:346–347
LazebnikY. A.,
TakahashiA.,
MoirR. D.,
GoldmanR. D.,
PoirierG. G.,
KaufmannS. H.,
EarnshawW. C.1995; Studies of the lamin proteinase reveal multiple parallel biochemical pathways during apoptotic execution. Proc Natl Acad Sci U S A 92:9042–9046
LiuX. S.,
ZouH.,
SlaughterC.,
WangX. D.1997; DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89:175–184
ManciniM.,
MachamerC. E.,
RoyS.,
NicholsonD. W.,
ThornberryN. A.,
Casciola-RosenL. A.,
RosenA.2000; Caspase-2 is localized at the Golgi complex and cleaves golgin-160 during apoptosis. J Cell Biol 149:603–612
NeillJ. D.1992; Nucleotide sequence of the capsid protein gene of 2 serotypes of San-Miguel sea lion virus – identification of conserved and nonconserved amino-acid-sequences among calicivirus capsid proteins. Virus Res 24:211–222
NeillJ. D.,
ReardonI. M.,
HeinriksonR. L.1991; Nucleotide sequence and expression of the capsid protein gene of feline calicivirus. J Virol 65:5440–5447
O'ReillyL. A.,
EkertP.,
HarveyN.11 other authors2002; Caspase-2 is not required for thymocyte or neuronal apoptosis even though cleavage of caspase-2 is dependent on both Apaf-1 and caspase-9. Cell Death Differ 9:832–841
OrthK.,
ChinnaiyanA. M.,
GargM.,
FroelichC. J.,
DixitV. M.1996; The CED-3/ICE-like protease Mch2 is activated during apoptosis and cleaves the death substrate lamin A. J Biol Chem 271:16443–16446
RobertsonJ. D.,
EnokssonM.,
SuomelaM.,
ZhivotovskyB.,
OrreniusS.2002; Caspase-2 acts upstream of mitochondria to promote cytochrome c release during etoposide-induced apoptosis. J Biol Chem 277:29803–29809
SamejimaK.,
SvingenP. A.,
BasiG. S.9 other authors1999; Caspase-mediated cleavage of DNA topoisomerase I at unconventional sites during apoptosis. J Biol Chem 274:4335–4340
SasakiS.,
XinK. Q.,
OkudelaK.,
OkudaK.,
IshiiN.2002; Immunomodulation by apoptosis-inducing caspases for an influenza DNA vaccine delivered by gene gun. Gene Ther 9:828–831
SleeE. A.,
AdrainC.,
MartinS. J.2001; Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis. J Biol Chem 276:7320–7326
SosnovtsevS. V.,
SosnovtsevaA. A.,
GreenK. Y.1998; Cleavage of the feline calicivirus capsid precursor is mediated by a virus-encoded proteinase. J Virol 72:3051–3059
SrinivasulaS. M.,
Fernandes-AlnemriT.,
ZangrilliJ.7 other authors1996; The Ced-3/interleukin 1 β converting enzyme-like homolog Mch6 and the lamin-cleaving enzyme Mch2 alpha are substrates for the apoptotic mediator CPP32. J Biol Chem 271:27099–27106
TakahashiA.,
AlnemriE. S.,
LazebnikY. A.7 other authors1996; Cleavage of lamin A by Mch2 alpha but not CPP32: multiple interleukin 1 β -converting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis. Proc Natl Acad Sci U S A 93:8395–8400
UbolS.,
KasisithJ.2000; Reactivation of Nedd-2, a develop-mentally down-regulated apoptotic gene, in apoptosis induced by a street strain of rabies virus. J Med Microbiol 49:1043–1046