Bioinformatic and experimental survey of 14-3-3-binding sites.
Bioinformatic and experimental survey of 14-3-3-binding sites.
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DOI：
10.1042/bj20091834
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发表时间：
2010-03-15
期刊：
影响因子：
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通讯作者：
MacKintosh C
中科院分区：
文献类型：
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作者：
Johnson C;Crowther S;Stafford MJ;Campbell DG;Toth R;MacKintosh C
分析了文献中200多个磷酸化14-3-3-结合位点，以定义14-3-3特异性，鉴定相关蛋白激酶，并深入了解细胞14-3-3/磷蛋白网络如何工作。模式I RXX（pS/pT）XP基序占主导地位，尽管+2脯氨酸残基出现在不到一半中，并且LX（R/K）SX（pS/pT）XP在植物14-3-3-结合位点中突出。脯氨酸在+1很少报道，这样的图案没有站起来的实验重新分析人类Ndel 1。相反，我们发现14-3-3与两个残基相互作用，这两个残基被嗜碱性激酶磷酸化，位于与认知障碍有关的Ndel 1的DISC 1（精神分裂症1中的破坏）相互作用区域。这些数据符合一般发现，即14-3-3-结合位点有不同的亚型，与不同嗜碱性AGC的特异性重叠（蛋白激酶A/蛋白激酶G/蛋白激酶C家族）和CaMK（Ca 2 +/钙调蛋白依赖性蛋白激酶）蛋白激酶，并且14-3-3二聚体通常与两个串联磷酸化位点接合，这是具有特殊信号传导的构型，机械和进化特性。因此，14-3-3二聚体可以是集成多个输入以产生动作的数字逻辑门，以及当两个结合位点被不同蛋白激酶磷酸化时的重合检测器。配对位点通常位于无序区域内和/或横跨功能结构域的任一侧，表明14-3-3二聚体如何调节其靶标的构象和/或相互作用。最后，14-3-3蛋白结合到几个多蛋白家族的成员。两个14-3-3-结合位点在IIa类组蛋白脱乙酰酶中是保守的，而其他蛋白质家族显示出由14-3-3s进行的差异调节。我们推测，14-3-3二聚体可能有助于这些家庭的进化，调整不同的生理需求的监管投入。
More than 200 phosphorylated 14-3-3-binding sites in the literature were analysed to define 14-3-3 specificities, identify relevant protein kinases, and give insights into how cellular 14-3-3/phosphoprotein networks work. Mode I RXX(pS/pT)XP motifs dominate, although the +2 proline residue occurs in less than half, and LX(R/K)SX(pS/pT)XP is prominent in plant 14-3-3-binding sites. Proline at +1 is rarely reported, and such motifs did not stand up to experimental reanalysis of human Ndel1. Instead, we discovered that 14-3-3 interacts with two residues that are phosphorylated by basophilic kinases and located in the DISC1 (disrupted-in-schizophrenia 1)-interacting region of Ndel1 that is implicated in cognitive disorders. These data conform with the general findings that there are different subtypes of 14-3-3-binding sites that overlap with the specificities of different basophilic AGC (protein kinase A/protein kinase G/protein kinase C family) and CaMK (Ca2+/calmodulin-dependent protein kinase) protein kinases, and a 14-3-3 dimer often engages with two tandem phosphorylated sites, which is a configuration with special signalling, mechanical and evolutionary properties. Thus 14-3-3 dimers can be digital logic gates that integrate more than one input to generate an action, and coincidence detectors when the two binding sites are phosphorylated by different protein kinases. Paired sites are generally located within disordered regions and/or straddle either side of functional domains, indicating how 14-3-3 dimers modulate the conformations and/or interactions of their targets. Finally, 14-3-3 proteins bind to members of several multi-protein families. Two 14-3-3-binding sites are conserved across the class IIa histone deacetylases, whereas other protein families display differential regulation by 14-3-3s. We speculate that 14-3-3 dimers may have contributed to the evolution of such families, tailoring regulatory inputs to different physiological demands.