Mitotic Substrates of the Kinase Aurora with Roles in Chromatin Regulation Identified Through Quantitative Phosphoproteomics of Fission Yeast
Mitotic Substrates of the Kinase Aurora with Roles in Chromatin Regulation Identified Through Quantitative Phosphoproteomics of Fission Yeast
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DOI：
10.1126/scisignal.2001588
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发表时间：
2011-06-28
影响因子：
7.3
通讯作者：
Hauf, Silke
中科院分区：
文献类型：
--
作者：
Koch, Andre;Krug, Karsten;Hauf, Silke
Aurora家族的激酶对于真核生物中有丝分裂的正确执行是必不可少的，并且Aurora抑制剂作为抗癌药物正在临床试验中。我们应用位点特异性定量磷酸化蛋白质组学结合化学抑制极光，以确定有丝分裂的极光基板在裂变酵母的蛋白质组范围内。我们检测到8000个磷酸化事件，其中我们将近6000个分配给特定的残基; 220个在暴露于Aurora抑制剂的细胞中减少。在控制了抑制剂的非特异性作用后，我们将70个位点（在42种蛋白质上）分类为Aurora的可能靶点，这使得Aurora磷酸化的共有序列得以细化。几种底物候选物是Aurora的已知靶点，验证了该方法，但大多数代表了新检测到的Aurora底物。这些极光基板在染色质动力学的各个方面的参与表明，除了其既定的作用，在控制染色体压实和附件的有丝分裂纺锤体，极光影响其他方面的染色质结构和功能在有丝分裂。
Kinases of the Aurora family are essential for the proper execution of mitosis in eukaryotes, and Aurora inhibitors are in clinical trials as anticancer drugs. We applied site-specific quantitative phosphoproteomics in conjunction with chemical inhibition of Aurora to identify mitotic Aurora substrates in fission yeast on a proteome-wide scale. We detected 8000 phosphorylation events, of which we assigned almost 6000 to a specific residue; 220 were reduced in cells exposed to the Aurora inhibitor. After controlling for unspecific effects of the inhibitor, we classified 70 sites (on 42 proteins) as probable targets of Aurora, which enabled refinement of the consensus sequence for phosphorylation by Aurora. Several of the substrate candidates were known targets of Aurora, validating the approach, but most represented newly detected Aurora substrates. The involvement of these Aurora substrates in diverse aspects of chromatin dynamics suggests that in addition to its established role in controlling chromosome compaction and attachment to the mitotic spindle, Aurora influences other aspects of chromatin architecture and function during mitosis.