Copper modulates the degradation of copper chaperone for Cu,Zn superoxide dismutase by the 26 S proteosome.
Copper chaperones are copper-binding proteins that directly insert copper into specific targets, preventing the accumulation of free copper ions that can be toxic to the cell. Despite considerable advances in the understanding of copper transfer from copper chaperones to their target, to date, there is no information regarding how the activity of these proteins is regulated in higher eukaryotes. The insertion of copper into the antioxidant enzyme Cu,Zn superoxide dismutase (SOD1) depends on the copper chaperone for SOD1 (CCS). We have recently reported that CCS protein is increased in tissues of rats fed copper-deficient diets suggesting that copper may regulate CCS expression. Here we show that whereas copper deficiency increased CCS protein in rats, mRNA level was unaffected. Rodent and human cell lines cultured in the presence of the specific copper chelator 2,3,2-tetraamine displayed a dose-dependent increase in CCS protein that could be reversed with the addition of copper but not iron or zinc to the cells. Switching cells from copper-deficient to copper-rich medium promoted the rapid degradation of CCS, which could be blocked by the proteosome inhibitors MG132 and lactacystin but not a cysteine protease inhibitor or inhibitors of the lysosomal degradation pathway. In addition, CCS degradation was slower in copper-deficient cells than in cells cultured in copper-rich medium. Together, these data show that copper regulates CCS expression by modulating its degradation by the 26 S proteosome and suggest a novel role for CCS in prioritizing the utilization of copper when it is scarce.
Bertinato
Jesse
J
Nutrition Research Division, Food Directorate, Health Products and Food Branch, Health Canada, 2203C Banting Research Centre, Ottawa, Ontario K1A 0L2, Canada.
L'Abbé
Mary R
MR
D000284:Administration, Oral; D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D003300:Copper / Q000008:administration & dosage / Q000172:deficiency / Q000494:pharmacology*; D019327:Copper Sulfate / Q000494:pharmacology; D017931:DNA Primers; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D008113:Liver Neoplasms; D008297:Male; D018832:Molecular Chaperones / Q000235:genetics*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011073:Polyamines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D021122:Protein Subunits / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D015183:Restriction Mapping; D013482:Superoxide Dismutase / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
C114678:CCS protein, human; C491961:Ccs protein, rat; D017931:DNA Primers; D018832:Molecular Chaperones; D011073:Polyamines; D011480:Protease Inhibitors; D021122:Protein Subunits; D012333:RNA, Messenger; C022177:2,3,2-tetramine; D003300:Copper; D013482:Superoxide Dismutase; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D019327:Copper Sulfate