Activity and expression of the 20S proteasome are increased in skeletal muscle during sepsis.
Recent studies suggest that sepsis stimulates ubiquitin-dependent protein breakdown in skeletal muscle. In this proteolytic pathway, ubiquitinated proteins are recognized, unfolded, and degraded by the multicatalytic 26S protease complex. The 20S proteasome is the catalytic core of the 26S protease complex. The role of the 20S proteasome in the regulation of sepsis-induced muscle proteolysis is not known. We tested the hypothesis that sepsis increases 20S proteasome activity and the expression of mRNA for various subunits of this complex. Proteolytic activity of isolated 20S proteasomes, assessed as activity against fluorogenic peptide substrates, was increased in extensor digitorum longus muscles from septic rats. The proteolytic activity was inhibited by specific proteasome blockers. Northern blot analysis revealed an approximately twofold increase in the relative abundance of mRNA for the 20S alpha-subunits RC3 and RC9 and the beta-subunit RC7. However, Western blot analysis did not show any difference in RC9 protein content between sham-operated and septic rats. The increased activity and expression of the 20S proteasome in muscles from septic rats lend further support for a role of the ubiquitin-proteasome-pathway in the regulation of sepsis-induced muscle proteolysis.
Hobler
S C
SC
Department of Surgery, Biochemistry and Microbiology, Howard Hughes Medical Institute, University of Cincinnati, 45267, USA.
Williams
A
A
Fischer
D
D
Wang
J J
JJ
Sun
X
X
Fischer
J E
JE
Monaco
J J
JJ
Hasselgren
P O
PO
D000818:Animals; D001424:Bacterial Infections / Q000378:metabolism*; D015152:Blotting, Northern; D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006614:Hindlimb; D008297:Male; D009097:Multienzyme Complexes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics; D010447:Peptide Hydrolases / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D009097:Multienzyme Complexes; D010446:Peptide Fragments; D011480:Protease Inhibitors; D012333:RNA, Messenger; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex