Human erythropoiesis is an exquisitely controlled multistep developmental process, and its dysregulation leads to numerous human diseases. Transcriptome and epigenome studies provided insights into system‐wide regulation, but we currently lack a global mechanistic view on the dynamics of proteome and post‐translational regulation coordinating erythroid maturation. We established a mass spectrometry (MS)‐based proteomics workflow to quantify and dynamically track 7,400 proteins and 27,000 phosphorylation sites of five distinct maturation stages of in vitro reconstituted erythropoiesis of CD34⁺ HSPCs. Our data reveal developmental regulation through drastic proteome remodeling across stages of erythroid maturation encompassing most protein classes. This includes various orchestrated changes in solute carriers indicating adjustments to altered metabolic requirements. To define the distinct proteome of each maturation stage, we developed a computational deconvolution approach which revealed stage‐specific marker proteins. The dynamic phosphoproteomes combined with a kinome‐targeted CRISPR/Cas9 screen uncovered coordinated networks of erythropoietic kinases and pinpointed downregulation of c‐Kit/MAPK signaling axis as key driver of maturation. Our system‐wide view establishes the functional dynamic of complex phosphosignaling networks and regulation through proteome remodeling in erythropoiesis.

中文翻译：

---

综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

人类红细胞生成是一个精确控制的多步骤发育过程，其失调会导致许多人类疾病。转录组和表观基因组研究提供了对全系统调节的见解，但我们目前缺乏对蛋白质组动力学和协调红系成熟的翻译后调节的全局机制观点。我们建立了基于质谱 (MS) 的蛋白质组学工作流程，以量化和动态跟踪CD34 ^{+ HSPC}体外重建红细胞生成的五个不同成熟阶段的 7,400 个蛋白质和 27,000 个磷酸化位点。我们的数据揭示了通过涵盖大多数蛋白质类别的红细胞成熟阶段的剧烈蛋白质组重塑来进行发育调节。这包括溶质载体的各种精心策划的变化，表明对改变的代谢需求的调整。为了定义每个成熟阶段的独特蛋白质组，我们开发了一种计算反卷积方法，该方法揭示了阶段特异性标记蛋白。动态磷酸化蛋白质组与激酶组靶向 CRISPR/Cas9 筛选相结合，发现了红细胞生成激酶的协调网络，并明确 c-Kit/MAPK 信号轴的下调是成熟的关键驱动因素。我们的全系统观点建立了复杂磷酸信号网络的功能动态以及通过红细胞生成过程中蛋白质组重塑的调节。