Sherpa Dawafuti, Mueller Judith, Karayel Özge, Xu Peng, Yao Yu, Chrustowicz Jakub, Gottemukkala Karthik V, Baumann Christine, Gross Annette, Czarnecki Oliver, Zhang Wei, Gu Jun, Nilvebrant Johan, Sidhu Sachdev S, Murray Peter J, Mann Matthias, Weiss Mitchell J, Schulman Brenda A, Alpi Arno F
Department of Molecular Machines and Signaling, Max Planck Institute of Biochemistry, Martinsried, Germany.
Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
Elife. 2022 Dec 2;11:e77937. doi: 10.7554/eLife.77937.
The development of haematopoietic stem cells into mature erythrocytes - erythropoiesis - is a controlled process characterized by cellular reorganization and drastic reshaping of the proteome landscape. Failure of ordered erythropoiesis is associated with anaemias and haematological malignancies. Although the ubiquitin system is a known crucial post-translational regulator in erythropoiesis, how the erythrocyte is reshaped by the ubiquitin system is poorly understood. By measuring the proteomic landscape of in vitro human erythropoiesis models, we found dynamic differential expression of subunits of the CTLH E3 ubiquitin ligase complex that formed maturation stage-dependent assemblies of topologically homologous RANBP9- and RANBP10-CTLH complexes. Moreover, protein abundance of CTLH's cognate E2 ubiquitin conjugating enzyme UBE2H increased during terminal differentiation, and UBE2H expression depended on catalytically active CTLH E3 complexes. CRISPR-Cas9-mediated inactivation of CTLH E3 assemblies or UBE2H in erythroid progenitors revealed defects, including spontaneous and accelerated erythroid maturation as well as inefficient enucleation. Thus, we propose that dynamic maturation stage-specific changes of UBE2H-CTLH E2-E3 modules control the orderly progression of human erythropoiesis.
造血干细胞发育为成熟红细胞的过程——红细胞生成——是一个受调控的过程，其特征在于细胞重组和蛋白质组格局的剧烈重塑。有序红细胞生成的失败与贫血和血液系统恶性肿瘤相关。尽管泛素系统是红细胞生成中已知的关键翻译后调节因子，但泛素系统如何重塑红细胞却知之甚少。通过测量体外人类红细胞生成模型的蛋白质组格局，我们发现CTLH E3泛素连接酶复合物亚基的动态差异表达，这些亚基形成了拓扑同源的RANBP9-和RANBP10-CTLH复合物的成熟阶段依赖性组装体。此外，CTLH同源E2泛素结合酶UBE2H的蛋白质丰度在终末分化期间增加，并且UBE2H的表达依赖于具有催化活性的CTLH E3复合物。CRISPR-Cas9介导的红细胞祖细胞中CTLH E3组装体或UBE2H的失活揭示了缺陷，包括自发和加速的红细胞成熟以及去核效率低下。因此，我们提出UBE2H-CTLH E2-E3模块的动态成熟阶段特异性变化控制着人类红细胞生成的有序进程。
