20S proteasomal degradation of ornithine decarboxylase is regulated by NQO1.
Ornithine decarboxylase (ODC), a key enzyme in the biosynthesis of polyamines, is a very labile protein. ODC is a homodimeric enzyme that undergoes ubiquitin-independent proteasomal degradation via direct interaction with antizyme, a polyamine-induced protein. Binding of antizyme promotes the dissociation of ODC homodimers and marks ODC for degradation by the 26S proteasomes. We describe here an alternative pathway for ODC degradation that is regulated by NAD(P)H quinone oxidoreductase 1 (NQO1). We show that NQO1 binds and stabilizes ODC. Dicoumarol, an inhibitor of NQO1, dissociates ODC-NQO1 interaction and enhances ubiquitin-independent ODC proteasomal degradation. We further show that dicoumarol sensitizes ODC monomers to proteasomal degradation in an antizyme-independent manner. This process of NQO1-regulated ODC degradation was recapitulated in vitro by using purified 20S proteasomes. Finally, we show that the regulation of ODC stability by NQO1 is especially prominent under oxidative stress. Our findings assign to NQO1 a role in regulating ubiquitin-independent degradation of ODC by the 20S proteasomes.
Asher
Gad
G
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
Bercovich
Zippi
Z
Tsvetkov
Peter
P
Shaul
Yosef
Y
Kahana
Chaim
C
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D001728:Dicumarol / Q000737:chemistry / Q000494:pharmacology; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000737:chemistry*; D009955:Ornithine Decarboxylase / Q000737:chemistry*; D018384:Oxidative Stress; D010957:Plasmids / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D013696:Temperature; D014162:Transfection; D025801:Ubiquitin / Q000737:chemistry
D025801:Ubiquitin; D001728:Dicumarol; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D046988:Proteasome Endopeptidase Complex; D009955:Ornithine Decarboxylase