Spermidine-induced destabilization of ornithine decarboxylase (ODC) is mediated by accumulation of antizyme in ODC-overproducing variant cells.
The mechanism of spermidine-induced destabilization of ornithine decarboxylase (ODC) was examined in newly isolated ODC-overproducing variant cells by use of an in vitro ODC degrading system. The cells accumulated ODC protein in the presence of alpha-difluoromethylornithine. Addition of spermidine to the medium accelerated degradation of ODC protein concomitantly with induction of antizyme, a regulatory protein that binds to ODC, inhibiting its activity. Both the acceleration of ODC degradation and the induction of antizyme were inhibited by cycloheximide, but not by actinomycin D. ODC was degraded rapidly in extracts from spermidine-treated cells. The rate of ODC degradation correlated with the amount of antizyme in the extracts, and the degradation activity was abolished by treatment of the extracts with anti-antizyme antibody. Thus, antizyme induced by spermidine was essential for the accelerated degradation of ODC in the cells. ODC was phosphorylated in the cells, probably at serine residue 303 in the first internal PEST region. ODC phosphorylation occurred even when its new synthesis was inhibited by cycloheximide. Antizyme accelerated the degradations of both dephosphorylated ODC and native ODC.
Kanamoto
R
R
Department of Nutrition, Jikei University School of Medicine, Tokyo, Japan.
Kameji
T
T
Iwashita
S
S
Igarashi
K
K
Hayashi
S
S
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002460:Cell Line; D002474:Cell-Free System; D002999:Clone Cells; D003545:Cysteine / Q000378:metabolism; D000518:Eflornithine / Q000494:pharmacology; D004790:Enzyme Induction; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D009955:Ornithine Decarboxylase / Q000096:biosynthesis / Q000378:metabolism*; D065108:Ornithine Decarboxylase Inhibitors; D011506:Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D013095:Spermidine / Q000494:pharmacology*; D013462:Sulfur Radioisotopes
D000906:Antibodies; D065108:Ornithine Decarboxylase Inhibitors; D011506:Proteins; D013462:Sulfur Radioisotopes; C031017:ornithine decarboxylase antizyme; D008715:Methionine; D009955:Ornithine Decarboxylase; D003545:Cysteine; D013095:Spermidine; D000518:Eflornithine