Involvement of the 20S proteasome in the degradation of ornithine decarboxylase

Abstract

Eukaryotic cells have been shown to contain two high-molecular-mass proteases of 700 kDa and 1400 kDa (20S and 26S proteases, respectively). It has been suggested that the 20S protease, also known as proteasome, may constitute the catalytic core of the 26S protease. While the role of the free 20S protease in intracellular protein degradation is unclear, the 26S protease is implicated in the degradation of ubiquinated proteins. We have recently demonstrated, that ornithine decarboxylase (ODC), one of the most labile proteins in mammalian cells, is degraded via an ATP-dependent but ubiquitin-independent proteolytic pathway. Here we extend these observations by demonstrating that in reticulocyte lysate ODC degradation is inhibited by antibodies raised against the C9 subunit of rat proteasome. Partial fractionation of the lysate demonstrated preferential degradation of ODC in the fraction of the lysate proteins that are precipitated by 38% ammonium sulfate. Since it was demonstrated that the 26S protease precipitates at this concentration of ammonium sulfate while the 20S proteasome remains soluble, our results suggest that the 26S protease is the one degrading ODC.