Proteolytic processing and assembly of the C5 subunit into the proteasome complex.
Assembly of mammalian 20 S proteasomes from individual subunits is beginning to be investigated. Proteasomes are made of four heptameric rings in the configuration alpha7beta7beta7alpha7. By using anti-proteasome and anti-subunit-specific antibodies, we characterized the processing and assembly of the beta subunit C5. The C5 precursor (25 kDa) remains as a free non-assembled polypeptide in the cell. The conversion of the C5 precursor to mature C5 (23 kDa) occurs concomitantly with its incorporation into 15 S proteasome intermediate and 20 S mature proteasome complexes. This processing is dependent on proteasome activity and takes place in the cytosol. These results are not fully compatible with the hypothesis that postulates that assembly of proteasomes takes place via a "half-proteasome" intermediate that contains one full alpha-ring and one full beta-ring of unprocessed beta subunit precursors.
Rodriguez-Vilariño
S
S
Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM, Facultad de Medicina de la Universidad Autónoma de Madrid, 28029 Madrid, Spain.
Arribas
J
J
Arizti
P
P
Castaño
J G
JG
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000276:immunology; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008099:Liver / Q000201:enzymology; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000276:immunology; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism
D000906:Antibodies; D009097:Multienzyme Complexes; D011498:Protein Precursors; D012333:RNA, Messenger; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex