Identification and characterization of a novel phosphoregulatory site on cyclin-dependent kinase 5.
Identification and characterization of a novel phosphoregulatory site on cyclin-dependent kinase 5.
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DOI：
10.1016/j.bbrc.2018.09.017
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发表时间：
2018-10-12
影响因子：
3.1
通讯作者：
Bhandari D
中科院分区：
文献类型：
--
作者：
Roach BL;Ngo JM;Limso C;Oloja KB;Bhandari D
细胞周期蛋白依赖性激酶5（CDK 5）是胚胎发育所必需的丝氨酸/苏氨酸激酶，其过度活化与包括神经变性、癌细胞转移和II型糖尿病在内的多种病理学有关。因此，重要的是研究介导CDK 5活性调节的分子机制。在这里，我们确定和表征一个新的磷酸化调节位点的CDK 5。我们的质谱分析确定了7个假定的磷酸化位点的CDK 5。使用拟磷酸化和非磷酸化突变体，我们确定，磷酸化的S47，一个确定的网站，使激酶催化失活。由于在S47处的磷酸化模拟变化导致的激酶的失活是由于其与其同源激活剂p35的相互作用的抑制。我们通过显示S47 D CDK 5突变体抑制细胞迁移并促进细胞增殖，将这种调节事件的作用与细胞表型联系起来。总之，这些结果揭示了调节CDK 5活性的潜在生理机制。不仅在CDK 5中，而且在大多数其他CDK家族成员中，可磷酸化残基（S/T）在该位置的进化位置表明，该磷酸化位点可能代表整个CDK家族的共享调节机制。
Cyclin-dependent kinase 5 (CDK5) is a serine/threonine kinase essential for embryonic development whose overactivation has been implicated in several pathologies including neurodegeneration, cancer cell metastasis and type II diabetes. Therefore, it is important to investigate molecular mechanism(s) that mediate regulation of CDK5 activity. Here we identify and characterize a novel phosphoregulatory site on CDK5. Our mass spectrometry analysis identified seven putative phosphorylation sites on CDK5. Using phosphomimetic and non-phosphorylatable mutants, we determined that phosphorylation of S47, one of the identified sites, renders the kinase catalytically inactive. The inactivation of the kinase due to the phosphomimetic change at S47 results from inhibition of its interaction with its cognate activator, p35. We connect the effect of this regulatory event to a cellular phenotype by showing that the S47D CDK5 mutant inhibits cell migration and promotes cell proliferation. Together, these results have uncovered a potential physiological mechanism to regulate CDK5 activity. The evolutionary placement of a phosphorylatable residue (S/T) at this position not only in CDK5 but also in the majority of other CDK family members suggests that this phosphosite may represent a shared regulatory mechanism across the CDK family.
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