Cdc20 proteolysis requires p38 MAPK signaling and Cdh1-independent APC/C ubiquitination during spindle assembly checkpoint activation by cadmium

• Published in: Journal of cellular physiology 2010-05, Vol.223 (2), p.327-334
• Main Authors: Yen, Ai-Hsin, Yang, Jia-Ling
• Format: Article
• Language: English
• Subjects: Anaphase-Promoting Complex-Cyclosome; Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; Cadherins - metabolism; Cadmium - pharmacology; Cdc20 Proteins; Cell Cycle Proteins - drug effects; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Line; Cell Line, Tumor; Cyclin A - drug effects; Cyclin A - metabolism; Enzyme Inhibitors - pharmacology; Genes, cdc - drug effects; Genes, cdc - physiology; Humans; MAP Kinase Kinase 6 - drug effects; MAP Kinase Kinase 6 - metabolism; MAP Kinase Signaling System - drug effects; MAP Kinase Signaling System - physiology; Mitogens - pharmacology; p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors; p38 Mitogen-Activated Protein Kinases - genetics; p38 Mitogen-Activated Protein Kinases - metabolism; Peptide Hydrolases - drug effects; Peptide Hydrolases - metabolism; Prometaphase - drug effects; Prometaphase - physiology; Spindle Apparatus - drug effects; Spindle Apparatus - metabolism; Ubiquitin-Protein Ligase Complexes - metabolism; Ubiquitination - drug effects; Ubiquitination - physiology
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.22038

Cdc20, an activator of the anaphase promoting complex/cyclosome (APC/C) ubiquitin ligase, initiates the destruction of key mitotic regulators to facilitate mitosis, while it is negatively regulated by the spindle assembly checkpoint (SAC) to prevent premature anaphase entry. Activation of the p38 mitogen‐activated protein kinase could contribute to mitotic arrest, but the underlying mechanism is unknown. Here we report a novel pathway in which the p38 signaling triggers Cdc20 destruction under SAC elicited by cadmium, a human carcinogen. We found that the cadmium‐induced prometaphase arrest was linked to decreased Cdc20 and accumulated cyclin A protein levels in human cells, whereas the activity of cyclin B1–Cdk1 was unaffected. The Cdc20 half‐life was markedly shortened along with its ubiquitination and degradation via 26S proteasome in cadmium‐treated asynchronous or G2‐enriched cells. Depletion of APC3 markedly suppressed the cadmium‐induced Cdc20 ubiquitination and proteolysis, while depletion of Cdh1, another activator of APC/C, did not. Intriguingly, blockage of p38 activity restored the Cdc20 levels for continuing mitosis under cadmium, while inhibition of JNK activity had no effect. The cadmium‐induced Cdc20 proteolysis was also suppressed during transient depletion of p38α or stable expression a dominant negative form of p38. Inhibition of p38 abolished the induction of Mad2–Cdc20–APC3 complex by cadmium. Moreover, forced expression of MKK6–p38 signaling could promote Cdc20 degradation in a Cdh1‐independent APC/C pathway. In summary, accelerated ubiquitination and proteolysis of Cdc20 is essential for prometaphase arrest that is mediated via the p38 signaling during SAC activation. J. Cell. Physiol. 223: 327–334, 2010. © 2010 Wiley‐Liss, Inc.