Quantitative Mass Spectrometry Analysis Reveals Similar Substrate Consensus Motif for Human Mps1 Kinase and Plk1

被引:65
作者
Dou, Zhen [1 ,3 ]
von Schubert, Conrad [2 ]
Koerner, Roman [1 ]
Santamaria, Anna [1 ,2 ]
Elowe, Sabine [1 ]
Nigg, Erich A. [1 ,2 ]
机构
[1] Max Planck Inst Biochem, Dept Cell Biol, D-82152 Martinsried, Germany
[2] Univ Basel, Biozentrum, Basel, Switzerland
[3] Univ Sci & Technol China, Hefei Natl Lab Phys Sci Microscale, Hefei 230026, Peoples R China
关键词
SPINDLE-ASSEMBLY CHECKPOINT; CENP-E; CHROMOSOMAL INSTABILITY; MITOTIC ARREST; PROTEIN-KINASE; PHOSPHORYLATION; ACTIVATION; LOCALIZATION; SPECIFICITY; EXPRESSION;
D O I
10.1371/journal.pone.0018793
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Members of the Mps1 kinase family play an essential and evolutionarily conserved role in the spindle assembly checkpoint (SAC), a surveillance mechanism that ensures accurate chromosome segregation during mitosis. Human Mps1 (hMps1) is highly phosphorylated during mitosis and many phosphorylation sites have been identified. However, the upstream kinases responsible for these phosphorylations are not presently known. Methodology/Principal Findings: Here, we identify 29 in vivo phosphorylation sites in hMps1. While in vivo analyses indicate that Aurora B and hMps1 activity are required for mitotic hyper-phosphorylation of hMps1, in vitro kinase assays show that Cdk1, MAPK, Plk1 and hMps1 itself can directly phosphorylate hMps1. Although Aurora B poorly phosphorylates hMps1 in vitro, it positively regulates the localization of Mps1 to kinetochores in vivo. Most importantly, quantitative mass spectrometry analysis demonstrates that at least 12 sites within hMps1 can be attributed to autophosphorylation. Remarkably, these hMps1 autophosphorylation sites closely resemble the consensus motif of Plk1, demonstrating that these two mitotic kinases share a similar substrate consensus. Conclusions/Significance: hMps1 kinase is regulated by Aurora B kinase and its autophosphorylation. Analysis on hMps1 autophosphorylation sites demonstrates that hMps1 has a substrate preference similar to Plk1 kinase.
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页数:9
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