Protein kinase CK2 and protein kinase D are associated with the COP9 signalosome
Protein kinase CK2 and protein kinase D are associated with the COP9 signalosome
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DOI：
10.1093/emboj/cdg127
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发表时间：
2003-03-17
期刊：
影响因子：
11.4
通讯作者：
Dubiel, W
中科院分区：
文献类型：
--
作者：
Uhle, S;Medalia, O;Dubiel, W
从人红细胞中纯化的COP 9信号体（CSN）具有磷酸化蛋白质如c-Jun和p53的激酶活性，从而导致其泛素（Ub）依赖性降解。在这里，我们显示蛋白激酶CK 2（CK 2）和蛋白激酶D（PKD）与CSN共纯化。免疫沉淀和远蛋白质印迹显示，CK 2和PKD实际上与CSN有关。如金标记ATP的电子显微镜所示，至少10%的CSN颗粒与激酶相关。激酶活性，最可能是由于CK 2和PKD，与来自HeLa细胞的CSN共免疫沉淀。CK 2与DeltaCSN 3（111-403）和CSN 7结合，而PKD与全长CSN 3相互作用。CK 2磷酸化CSN 2和CSN 7，PKD修饰CSN 7。CK 2和PKD都磷酸化c-Jun以及p53。CK 2磷酸化Thr 155，其靶向p53以通过Ub系统降解。姜黄素、大黄素、DRB和白藜芦醇阻断CSN相关激酶并诱导HeLa细胞中c-Jun降解。姜黄素治疗导致c-Jun-Ub缀合物的量升高。我们的结论是，CK 2和PKD招募CSN，以调节Ub结合物的形成。
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. Immunoprecipi tation 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.