Phosphorylation of CLIP-170 by Plk1 and CK2 promotes timely formation of kinetochore-microtubule attachments
Phosphorylation of CLIP-170 by Plk1 and CK2 promotes timely formation of kinetochore-microtubule attachments
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DOI：
10.1038/emboj.2010.174
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发表时间：
2010-09-01
期刊：
影响因子：
11.4
通讯作者：
Liu, Xiaoqi
中科院分区：
文献类型：
--
作者：
Li, Hongchang;Liu, X. Shawn;Liu, Xiaoqi
CLIP-170通过与动力蛋白/动力肌动蛋白复合物直接相互作用而参与运动舞蹈-微管附着的形成。然而，CLIP-170的这一重要功能是否受到磷酸化的调节尚不清楚。在此，我们已经确定了Polo样激酶1（Plk 1）和酪蛋白激酶2（CK 2）作为CLIP-170的两种激酶，并将S195和S1318定位为它们各自的磷酸化位点。我们发现，CK 2 unphosphorylatable突变体失去了结合dynactin的能力，并在前中期定位到动粒，这表明CK 2磷酸化的CLIP-170参与其dynactin介导的动粒定位。此外，我们提供的证据表明，Plk 1磷酸化的CLIP-170在S195增强其与CK 2。最后，我们检测到缺陷的细胞中表达的CLIP-S195 A和-S1318 A，但不是CLIP-S195 E和-S1318 D的动粒纤维的形成，证实PLK 1和CK 2相关的磷酸化的CLIP-170参与有丝分裂中的动粒微管附件的及时形成。The EMBO Journal（2010）29，2953-2965. doi：10.1038/daj.2010.174; 2010年7月27日在线发布
CLIP-170 is implicated in the formation of kinetochore-microtubule attachments through direct interaction with the dynein/dynactin complex. However, whether this important function of CLIP-170 is regulated by phosphorylation is unknown. Herein, we have identified polo-like kinase 1 (Plk1) and casein kinase 2 (CK2) as two kinases of CLIP-170 and mapped S195 and S1318 as their respective phosphorylation sites. We showed that a CK2 unphosphorylatable mutant lost its ability to bind to dynactin and to localize to kinetochores during prometaphase, indicating that the CK2 phosphorylation of CLIP-170 is involved in its dynactin-mediated kinetochore localization. Furthermore, we provide evidence that Plk1 phosphorylation of CLIP-170 at S195 enhances its association with CK2. Finally, we detected defects in the formation of kinetochore fibres in cells expressing the CLIP-S195A and -S1318A, but not the CLIP-S195E and -S1318D, confirming that Plk1-and CK2-associated phosphorylations of CLIP-170 are involved in the timely formation of kinetochore-microtubule attachments in mitosis. The EMBO Journal (2010) 29, 2953-2965. doi: 10.1038/emboj.2010.174; Published online 27 July 2010