Molecular characteristics of porcine Fas-associated death domain (FADD) and procaspase-8.
To reveal the intracellular signal transduction molecules involved in granulosa cell apoptosis in porcine ovarian follicles, we cloned the porcine Fas-associated death domain (FADD), an adaptor protein for the cell death receptor, and procaspase-8, an initiator caspase. Porcine FADD (pFADD) was 636 bp (211 amino acids: aa) long and showed 74.0 and 65.4% homology with human and murine FADD, respectively. Porcine procaspase-8 (pprocaspase-8) was 1,431 bp (476 aa) long and 70.6 and 63.4% homologous with human and murine procaspase-8, respectively. To confirm the apoptosis-inducing abilities, we constructed pFADD and pprocaspase-8 cDNA expression vectors with enhanced green fluorescence protein (EGFP) and then transfected them into human uterine cervix tumor (HeLa-K), human granulosa cell-derived (KGN), murine granulosa-derived tumor (KK1), and porcine granulosa cell-derived (JC410) cells. When pFADD and pprocaspase-8 were overexpressed, cell death was induced in these transfected cells. However when caspase-inhibitor p35 was cotransfected, cell death was inhibited. The pFADD and pprocaspase-8 genes are well conserved, as are the physiological functions of their products.
Inoue
Naoko
N
Laboratory of Animal Morphology and Function, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
Matsuda-Minehata
Fuko
F
Goto
Yasufumi
Y
Sakamaki
Kazuhiro
K
Manabe
Noboru
N
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D053181:Caspase 8 / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D053200:Fas-Associated Death Domain Protein / Q000235:genetics* / Q000378:metabolism; D005260:Female; D006107:Granulosa Cells / Q000473:pathology / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000235:genetics*
D053200:Fas-Associated Death Domain Protein; D053181:Caspase 8