Protein kinase CK2 and protein kinase D are associated with the COP9 signalosome.
The COP9 signalosome (CSN) purified from human erythrocytes possesses kinase activity that phosphoryl ates proteins such as c-Jun and p53 with consequence for their ubiquitin (Ub)-dependent degradation. Here we show that protein kinase CK2 (CK2) and protein kinase D (PKD) co-purify with CSN. Immunoprecipitation and far-western blots reveal that CK2 and PKD are in fact associated with CSN. As indicated by electron microscopy with gold-labeled ATP, at least 10% of CSN particles are associated with kinases. Kinase activity, most likely due to CK2 and PKD, co-immuno precipitates with CSN from HeLa cells. CK2 binds to DeltaCSN3(111-403) and CSN7, whereas PKD interacts with full-length CSN3. CK2 phosphorylates CSN2 and CSN7, and PKD modifies CSN7. Both CK2 and PKD phosphorylate c-Jun as well as p53. CK2 phosphoryl ates Thr155, which targets p53 to degradation by the Ub system. Curcumin, emodin, DRB and resveratrol block CSN-associated kinases and induce degradation of c-Jun in HeLa cells. Curcumin treatment results in elevated amounts of c-Jun-Ub conjugates. We conclude that CK2 and PKD are recruited by CSN in order to regulate Ub conjugate formation.
Uhle
Stefan
S
Division of Molecular Biology, Department of Surgery, Institute of Toxicology, Medical Faculty Charité, Humboldt University, Monbijoustrasse 2, D-10117 Berlin, Germany.
Medalia
Ohad
O
Waldron
Richard
R
Dumdey
Renate
R
Henklein
Peter
P
Bech-Otschir
Dawadschargal
D
Huang
Xiaohua
X
Berse
Matthias
M
Sperling
Joseph
J
Schade
Rüdiger
R
Dubiel
Wolfgang
W
D000255:Adenosine Triphosphate / Q000378:metabolism; D000075686:COP9 Signalosome Complex; D047390:Casein Kinase II; D003474:Curcumin / Q000494:pharmacology; D004268:DNA-Binding Proteins; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D004642:Emodin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004912:Erythrocytes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D046912:Multiprotein Complexes; D010447:Peptide Hydrolases; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D021122:Protein Subunits; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D000077185:Resveratrol; D015398:Signal Transduction; D013267:Stilbenes / Q000494:pharmacology; D013379:Substrate Specificity; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D014452:Ubiquitins / Q000378:metabolism
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D046912:Multiprotein Complexes; D021122:Protein Subunits; D011506:Proteins; D016755:Proto-Oncogene Proteins c-jun; D011994:Recombinant Proteins; D013267:Stilbenes; D016159:Tumor Suppressor Protein p53; D014452:Ubiquitins; D004004:Dichlororibofuranosylbenzimidazole; D000255:Adenosine Triphosphate; C089212:protein kinase D; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C; D010447:Peptide Hydrolases; D000075686:COP9 Signalosome Complex; D003474:Curcumin; D004642:Emodin; D000077185:Resveratrol