Faustova Ilona, Örd Mihkel, Kiselev Viacheslav, Fedorenko Dmytro, Borovko Irina, Macs Dags, Pääbo Kaur, Lõoke Marko, Loog Mart
Institute of Technology, University of Tartu, Tartu 50411, Estonia.
Sci Adv. 2022 Aug 19;8(33):eabp8992. doi: 10.1126/sciadv.abp8992. Epub 2022 Aug 17.
The complexity of multisite phosphorylation mechanisms in regulating nuclear localization signals (NLSs) and nuclear export signals (NESs) is not understood, and its potential has not been used in synthetic biology. The nucleocytoplasmic shuttling of many proteins is regulated by cyclin-dependent kinases (CDKs) that rely on multisite phosphorylation patterns and short linear motifs (SLiMs) to dynamically control proteins in the cell cycle. We studied the role of motif patterns in nucleocytoplasmic shuttling using sensors based on the CDK targets Dna2, Psy4, and Mcm2/3 of . We designed multisite phosphorylation modules by rearranging phosphorylation sites, cyclin-specific SLiMs, phospho-priming, phosphatase specificity, and NLS/NES phospho-regulation and obtained very different substrate localization dynamics. These included ultrasensitive responses with and without a delay, graded responses, and different homeostatic plateaus. Thus, CDK can do much more than trigger sequential switches during the cell cycle as it can drive complex patterns of protein localization and activity by using multisite phosphorylation networks.
多位点磷酸化机制在调节核定位信号（NLSs）和核输出信号（NESs）方面的复杂性尚未被理解，并且其潜力尚未在合成生物学中得到应用。许多蛋白质的核质穿梭由细胞周期蛋白依赖性激酶（CDKs）调节，这些激酶依赖多位点磷酸化模式和短线性基序（SLiMs）来在细胞周期中动态控制蛋白质。我们使用基于CDK靶点Dna2、Psy4和Mcm2/3的传感器研究了基序模式在核质穿梭中的作用。我们通过重新排列磷酸化位点、细胞周期蛋白特异性SLiMs、磷酸化引发、磷酸酶特异性以及NLS/NES磷酸化调节来设计多位点磷酸化模块，并获得了非常不同的底物定位动态。这些包括有延迟和无延迟的超敏反应、分级反应以及不同的稳态平台。因此，CDK在细胞周期中所做的远不止触发顺序开关，因为它可以通过使用多位点磷酸化网络驱动复杂的蛋白质定位和活性模式。
