Destabilization of ornithine decarboxylase by transfected antizyme gene expression in hepatoma tissue culture cells.
The degradation of ornithine decarboxylase (ODC) is stimulated by polyamines in a protein synthesis-dependent manner. It has been suggested that antizyme, an ODC-inhibiting protein induced by polyamines, is involved in the process of polyamine-stimulated ODC decay. In this study, we investigated the direct effect of antizyme on ODC decay in hepatoma tissue culture (HTC) cells. A truncated rat antizyme cDNA, Z1, was inserted into an expression vector at a site under the control of a glucocorticoid-inducible promoter and transfected into HTC cells. In the transfected cells dexamethasone increased the amount of Z1 mRNA and induced active antizyme in the absence of exogenous polyamines. When dexamethasone was added to cells with a high level of ODC, rapid decays of ODC activity and protein were elicited after a lag time. Cycloheximide abolished the effect of dexamethasone. These effects of dexamethasone were not observed in control HTC cells transfected with the chloramphenicol acetyltransferase gene. This study indicated that, once induced, antizyme stimulated ODC degradation independently of polyamines and strongly supported our previous hypothesis that the ODC decay-accelerating action of polyamines is mediated by antizyme.
Murakami
Y
Y
Department of Nutrition, Jikei University School of Medicine, Tokyo, Japan.
Matsufuji
S
S
Miyazaki
Y
Y
Hayashi
S
S
D000818:Animals; D015152:Blotting, Northern; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D009955:Ornithine Decarboxylase / Q000378:metabolism*; D065108:Ornithine Decarboxylase Inhibitors; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
D065108:Ornithine Decarboxylase Inhibitors; D012333:RNA, Messenger; D003907:Dexamethasone; D004247:DNA; D003513:Cycloheximide; D015500:Chloramphenicol O-Acetyltransferase; D009955:Ornithine Decarboxylase