Porcine caspase-3: its cloning and activity during apoptosis of PK15 cells induced by porcine Fas ligand.
We cloned and sequenced cDNA that contained the coding sequence of porcine caspase-3. The open reading frame (ORF) of porcine caspase-3 cDNA was 834 base pairs (bp) in length and encoded 277 amino acids. The predicted amino acid sequence was 88.4%, 86.6%, and 87.7% homologous to the predicted human, murine, and rat amino acid sequences, respectively. The activity of caspase-3 in porcine renal tubular cell line PK15 after recombinant porcine Fas ligand (FasL) stimulation was examined. The enzymatic activity of caspase-3, but not that of caspase-1, was significantly increased after FasL treatment. Western blot analysis also showed that the processing of caspase-3 from proenzyme to mature subunits occurred after FasL treatment. The inhibition of caspase-3 by its specific inhibitor partially prevented the apoptotic cell death of PK15 cells caused by FasL. The porcine caspase-3 cDNA isolated in this study will be useful for the study of apoptotic cell death in pigs and will lead to the discovery of therapeutic uses of caspases and their inhibitors in the prevention of viral and bacterial diseases and tissue injury associated with xenotransplantation and allotransplantation.
Muneta
Y
Y
Institute of Animal Health, Tsukuba, Ibaraki, 305-0856, Japan. ymuneta@niah.affrc.go.jp
Shimoji
Y
Y
Mori
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013552:Swine
D061945:Caspase Inhibitors; D017931:DNA Primers; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003513:Cycloheximide; C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases