Klemm Cinzia, Ólafsson Guðjón, Wood Henry Richard, Mellor Caitlin, Zabet Nicolae Radu, Thorpe Peter Harold
School of Biological and Behavioural Sciences, Queen Mary University of London, London, UK.
Department of Bioengineering, Imperial College London, London, UK.
Nucleus. 2024 Dec;15(1):2420129. doi: 10.1080/19491034.2024.2420129. Epub 2024 Dec 1.
Dynamic protein phosphorylation and dephosphorylation play an essential role in cell cycle progression. Kinases and phosphatases are generally highly conserved across eukaryotes, underlining their importance for post-translational regulation of substrate proteins. In recent years, advances in phospho-proteomics have shed light on protein phosphorylation dynamics throughout the cell cycle, and ongoing progress in bioinformatics has significantly improved annotation of specific phosphorylation events to a given kinase. However, the functional impact of individual phosphorylation events on cell cycle progression is often unclear. To address this question, we used the Synthetic Physical Interactions (SPI) method, which enables the systematic recruitment of phospho-regulators to most yeast proteins. Using this method, we identified several putative novel targets involved in chromosome segregation and cytokinesis. The SPI method monitors cell growth and, therefore, serves as a tool to determine the impact of protein phosphorylation on cell cycle progression.
动态蛋白质磷酸化和去磷酸化在细胞周期进程中起着至关重要的作用。激酶和磷酸酶在真核生物中通常高度保守，这凸显了它们对底物蛋白翻译后调控的重要性。近年来，磷酸蛋白质组学的进展揭示了整个细胞周期中的蛋白质磷酸化动态，并且生物信息学的不断进步显著改善了对特定激酶磷酸化事件的注释。然而，单个磷酸化事件对细胞周期进程的功能影响往往并不清楚。为了解决这个问题，我们使用了合成物理相互作用（SPI）方法，该方法能够将磷酸调节因子系统地募集到大多数酵母蛋白上。通过使用这种方法，我们鉴定出了几个参与染色体分离和胞质分裂的假定新靶点。SPI方法监测细胞生长，因此可作为确定蛋白质磷酸化对细胞周期进程影响的工具。
