Human erythropoiesis is exquisitely controlled at multiple levels and its dysregulation leads to numerous human diseases. Despite many functional studies focused on classical regulators, we lack a global, system-wide understanding of post-translational mechanisms coordinating erythroid maturation. Using the latest advances in mass spectrometry (MS)-based proteomics we comprehensively investigate the dynamics of protein and post-translational regulation of in vitro reconstituted CD34+ HSPC-derived erythropoiesis. This quantifies and dynamically tracks 7,400 proteins and 27,000 phosphorylation sites. Our data reveals differential temporal protein expression encompassing most protein classes and numerous post-translational regulatory cascades. Drastic cell surface remodeling across erythropoiesis include numerous orchestrated changes in solute carriers, providing new stage-specific markers. The dynamic phosphoproteomes combined with a kinome-targeting CRISPR/Cas9 screen reveal coordinated networks of erythropoietic kinases and downregulation of MAPK signaling subsequent to c-Kit attenuation as key drivers of maturation. Our global view of erythropoiesis establishes a central role of post-translational regulation in terminal differentiation.

中文翻译：

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整合蛋白质组学揭示了人类红细胞生成中动态磷酸信号网络的原理

人类的红细胞生成被精确地控制在多个水平，其失调导致许多人类疾病。尽管进行了许多针对经典调节剂的功能研究，但我们对协调红细胞成熟的翻译后机制缺乏全局性的系统了解。利用基于质谱（MS）的蛋白质组学的最新进展，我们全面研究了蛋白质的动力学和体外重组CD34 + HSPC衍生的红细胞生成的翻译后调控。这将量化并动态跟踪7,400个蛋白质和27,000个磷酸化位点。我们的数据揭示了差异性的时间蛋白表达，其涵盖了大多数蛋白类别和许多翻译后调节级联。整个红细胞生成过程中的剧烈细胞表面重塑包括溶质载体的许多精心策划的变化，从而提供了新的阶段特异性标记。动态磷酸化蛋白质组与靶向激酶组的CRISPR / Cas9筛查相结合，揭示了促红细胞生成的激酶的协调网络和c-Kit衰减后MAPK信号的下调作为成熟的关键驱动力。我们对红细胞生成的整体看法确立了翻译后调控在终末分化中的核心作用。