A phospho-proteomic screen identifies substrates of the checkpoint kinase Chk1.
A phospho-proteomic screen identifies substrates of the checkpoint kinase Chk1.
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DOI：
10.1186/gb-2011-12-8-r78
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发表时间：
2011-08-18
期刊：
影响因子：
12.3
通讯作者：
Jackson SP
中科院分区：
文献类型：
--
作者：
Blasius M;Forment JV;Thakkar N;Wagner SA;Choudhary C;Jackson SP
细胞周期检查点激酶Chk1在哺乳动物细胞中是必不可少的，因为它在控制DNA复制、有丝分裂和DNA损伤反应等过程中起作用。尽管其至关重要，Chk1如何控制这些功能仍不清楚，主要是因为很少有Chk1基板迄今已被确定。在这里，我们结合联合收割机的化学遗传学方法与高分辨率质谱法，以确定新的Chk1底物及其磷酸化位点。产生的靶标列表揭示了Chk1功能的潜在影响，不仅对已知Chk1参与的过程，而且对其他关键细胞事件，如转录，RNA剪接和细胞命运决定。此外，我们验证和探讨磷酸化的转录辅阻遏物KAP1 Ser473作为一种新的DNA损伤诱导的Chk1网站。通过提供大量的潜在Chk1底物，我们提出了机会，研究Chk1在控制广泛的细胞过程中的意想不到的功能。我们还完善了Chk1的共识序列，促进未来的预测Chk1的目标网站。此外，我们鉴定的KAP1 Ser473磷酸化作为Chk1活性的稳健读数，可用于探索正在开发用于临床评价的Chk1抑制剂的体内作用。
The cell-cycle checkpoint kinase Chk1 is essential in mammalian cells due to its roles in controlling processes such as DNA replication, mitosis and DNA-damage responses. Despite its paramount importance, how Chk1 controls these functions remains unclear, mainly because very few Chk1 substrates have hitherto been identified. Here, we combine a chemical genetics approach with high-resolution mass spectrometry to identify novel Chk1 substrates and their phosphorylation sites. The list of targets produced reveals the potential impact of Chk1 function not only on processes where Chk1 was already known to be involved, but also on other key cellular events such as transcription, RNA splicing and cell fate determination. In addition, we validate and explore the phosphorylation of transcriptional co-repressor KAP1 Ser473 as a novel DNA-damage-induced Chk1 site. By providing a substantial set of potential Chk1 substrates, we present opportunities for studying unanticipated functions for Chk1 in controlling a wide range of cellular processes. We also refine the Chk1 consensus sequence, facilitating the future prediction of Chk1 target sites. In addition, our identification of KAP1 Ser473 phosphorylation as a robust readout for Chk1 activity could be used to explore the in vivo effects of Chk1 inhibitors that are being developed for clinical evaluation.
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DOI：
10.1083/jcb.200905059
发表时间：
2010-03-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Beck H;Nähse V;Larsen MS;Groth P;Clancy T;Lees M;Jørgensen M;Helleday T;Syljuåsen RG;Sørensen CS
通讯作者：
Sørensen CS
影响因子：
21.3
作者：
Jazayeri, A;Falck, J;Jackson, SP
通讯作者：
Jackson, SP
影响因子：
4.3
作者：
Goloudina, Anastasia;Yamaguchi, Hiroshi;Bulavin, Dmitry V.
通讯作者：
Bulavin, Dmitry V.
DOI：
10.1073/pnas.0708966105
发表时间：
2008-02-05
影响因子：
11.1
作者：
Blethrow, Justin D.;Glavy, Joseph S.;Shokat, Kevan M.
通讯作者：
Shokat, Kevan M.
影响因子：
7
作者：
Bennetzen, Martin V.;Larsen, Dorthe Helena;Andersen, Jens S.
通讯作者：
Andersen, Jens S.