Quantitative mass spectrometry analysis reveals similar substrate consensus motif for human Mps1 kinase and Plk1.
Quantitative mass spectrometry analysis reveals similar substrate consensus motif for human Mps1 kinase and Plk1.
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DOI：
10.1371/journal.pone.0018793
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发表时间：
2011-04-13
期刊：
影响因子：
3.7
通讯作者：
Nigg EA
中科院分区：
文献类型：
--
作者：
Dou Z;von Schubert C;Körner R;Santamaria A;Elowe S;Nigg EA
Mps1激酶家族成员在纺锤体组装检查点（SAC）中发挥着重要的进化保守作用，这是一种确保有丝分裂期间染色体准确分离的监视机制。人类Mps1 （hMps1）在有丝分裂过程中被高度磷酸化，许多磷酸化位点已经被确定。然而，负责这些磷酸化的上游激酶目前尚不清楚。在这里，我们在hMps1中鉴定了29个体内磷酸化位点。虽然体内分析表明，hMps1的有丝分裂超磷酸化需要Aurora B和hMps1的活性，但体外激酶实验表明，Cdk1、MAPK、Plk1和hMps1本身可以直接磷酸化hMps1。尽管在体外，Aurora B对hMps1的磷酸化作用较弱，但在体内，它能积极调节Mps1向着丝点的定位。最重要的是，定量质谱分析表明，hMps1中至少有12个位点可归因于自磷酸化。值得注意的是，这些hMps1自磷酸化位点与Plk1的共识基序非常相似，表明这两种有丝分裂激酶具有相似的底物共识。hMps1激酶受Aurora B激酶及其自磷酸化调控。对hMps1自磷酸化位点的分析表明，hMps1具有类似于Plk1激酶的底物偏好。
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. 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. 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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DOI：
10.1038/nrm2718
发表时间：
2009-07
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.8
作者：
Mattison, Christopher P.;Old, William M.;Winey, Mark
通讯作者：
Winey, Mark
影响因子：
10.5
作者：
Elowe, Sabine;Huemmer, Stefan;Nigg, Erich A.
通讯作者：
Nigg, Erich A.
影响因子：
3.3
作者：
Liu, ST;Chan, GKT;Yen, TJ
通讯作者：
Yen, TJ
DOI：
10.1083/jcb.200910027
发表时间：
2010-04-05
期刊：
The Journal of cell biology
影响因子：
--
作者：
Araki Y;Gombos L;Migueleti SP;Sivashanmugam L;Antony C;Schiebel E
通讯作者：
Schiebel E