Identification of DYRK1B as a substrate of ERK1/2 and characterisation of the kinase activity of DYRK1B mutants from cancer and metabolic syndrome.
Identification of DYRK1B as a substrate of ERK1/2 and characterisation of the kinase activity of DYRK1B mutants from cancer and metabolic syndrome.
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DOI：
10.1007/s00018-015-2032-x
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发表时间：
2016-02
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影响因子：
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通讯作者：
Cook SJ
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作者：
Ashford AL;Dunkley TP;Cockerill M;Rowlinson RA;Baak LM;Gallo R;Balmanno K;Goodwin LM;Ward RA;Lochhead PA;Guichard S;Hudson K;Cook SJ
双特异性酪氨酸磷酸化调节激酶DYRK 1B在肌生成过程中重新表达，在某些癌症中扩增或突变，并在代谢综合征的家族性病例中突变。DYRK 1B在翻译过程中通过激活环内酪氨酸-273（Y273）上的顺式自磷酸化来激活，但迄今为止很少有其他DYRK 1B磷酸化位点被表征。在这里，我们证明，DYRK 1B也经历了丝氨酸-421（S421）在体外和细胞中的反式自磷酸化，这个网站有助于DYRK 1B激酶活性。虽然DYRK 1BS 421 A突变体在细胞中完全缺乏p-S421，但DYRK 1B抑制剂仅引起p-S421的部分损失，表明存在也可以磷酸化DYRK 1B S421的额外激酶。事实上，无催化活性的DYRK 1BD 239 A突变体在细胞中表现出非常低的p-S421水平，但这被KRASG 12 V增加。此外，RAF-MEK 1/2-ERK 1/2信号通路的选择性激活迅速增加细胞中的p-S421，而应激激酶JNK或p38的激活则不能。S421位于Ser-Pro磷酸受体基序内，该基序是体外ERK 1/2和重组ERK 2在S421处磷酸化DYRK 1B的典型基序。我们的研究结果表明，DYRK 1B是一种新的ERK 2底物，揭示了参与细胞命运决定的两种激酶之间的新联系。最后，我们表明，DYRK 1B突变体，最近被描述在癌症和代谢综合征表现出正常或降低的内在激酶活性。本文的在线版本（doi：10.1007/s 00018 -015-2032-x）包含补充材料，可供授权用户使用。
The dual-specificity tyrosine-phosphorylation-regulated kinase, DYRK1B, is expressed de novo during myogenesis, amplified or mutated in certain cancers and mutated in familial cases of metabolic syndrome. DYRK1B is activated by cis auto-phosphorylation on tyrosine-273 (Y273) within the activation loop during translation but few other DYRK1B phosphorylation sites have been characterised to date. Here, we demonstrate that DYRK1B also undergoes trans-autophosphorylation on serine-421 (S421) in vitro and in cells and that this site contributes to DYRK1B kinase activity. Whilst a DYRK1BS421A mutant was completely defective for p-S421 in cells, DYRK1B inhibitors caused only a partial loss of p-S421 suggesting the existence of an additional kinase that could also phosphorylate DYRK1B S421. Indeed, a catalytically inactive DYRK1BD239A mutant exhibited very low levels of p-S421 in cells but this was increased by KRASG12V. In addition, selective activation of the RAF-MEK1/2-ERK1/2 signalling pathway rapidly increased p-S421 in cells whereas activation of the stress kinases JNK or p38 could not. S421 resides within a Ser-Pro phosphoacceptor motif that is typical for ERK1/2 and recombinant ERK2 phosphorylated DYRK1B at S421 in vitro. Our results show that DYRK1B is a novel ERK2 substrate, uncovering new links between two kinases involved in cell fate decisions. Finally, we show that DYRK1B mutants that have recently been described in cancer and metabolic syndrome exhibit normal or reduced intrinsic kinase activity. The online version of this article (doi:10.1007/s00018-015-2032-x) contains supplementary material, which is available to authorized users.