The ubiquitin-proteasome system is responsible for cysteine-responsive regulation of cysteine dioxygenase concentration in liver.
Hepatic cysteine dioxygenase (CDO) activity is a critical regulator of cellular cysteine concentration and availability of cysteine for anabolic processes and is markedly higher in animals fed diets containing excess sulfur amino acids compared with those fed levels at or below the requirement. Rat hepatocytes responded to a deficiency or excess of cysteine in the culture medium with a decrease or increase in CDO level but no change in CDO mRNA level. The cysteine analog, cysteamine, but not cysteine metabolites or thiol reagents, was also effective in increasing CDO. Inhibitors of the 26S proteasome blocked CDO degradation in cysteine-deficient cells but had little or no effect on CDO concentration in hepatocytes cultured with excess cysteine. High-molecular-mass CDO-ubiquitin conjugates were observed in cells cultured in cysteine-deficient medium, whether or not proteasome inhibitor was present, but these CDO-ubiquitin conjugates were not observed in cells cultured in cysteine-supplemented medium with or without proteasome inhibitor. Similar results were observed for degradation of recombinant CDO expressed in human heptocarcinoma cells cultured in cysteine-deficient or cysteine-supplemented medium. CDO is an example of a mammalian enzyme that is robustly regulated via its substrate, with the presence of substrate blocking the ubiquitination of CDO and, hence, the targeting of CDO for proteasomal degradation. This regulation occurs in primary hepatocytes in a manner that corresponds with changes observed in intact animals.
Stipanuk
Martha H
MH
Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853-6301, USA. mhs6@cornell.edu
Hirschberger
Lawrence L
LL
Londono
Monica P
MP
Cresenzi
Carrie L
CL
Yu
Anthony F
AF
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism*; D050558:Cysteine Dioxygenase; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D049308:Dioxygenases; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000378:metabolism*; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008297:Male; D009097:Multienzyme Complexes / Q000378:metabolism*; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D051381:Rats; D017207:Rats, Sprague-Dawley; D025801:Ubiquitin / Q000378:metabolism*
D009097:Multienzyme Complexes; D025801:Ubiquitin; D010105:Oxygenases; D049308:Dioxygenases; D050558:Cysteine Dioxygenase; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D003545:Cysteine