The ubiquitination or SUMOylation of hematopoietic related factors plays pivotal roles in hematopoiesis. RNF111, known as a ubiquitin ligase (Ubl), is a newly discovered SUMO-targeted ubiquitin ligase (STUbl) involved in multiple signaling pathways mediated by TGF-β family members. However, its role in hematopoiesis remains unclear. Herein, a heritable Rnf111 mutant zebrafish line was generated by CRISPR/Cas9-mediated genome editing. Impaired hematopoietic stem and progenitor cells (HSPC) of definitive hematopoiesis was found in Rnf111 deficient mutants. Ablation of Rnf111 resulted in decreased phosphorylation of Smad2/3 in HSPC. Definitive endoderm 2 inducer (IDE2), which specifically activates TGF-β signaling and downstream Smad2 phosphorylation, can restore the definitive hematopoiesis in Rnf111-deficient embryos. Further molecular mechanism studies revealed that Gcsfr/NO signaling was an important target pathway of Smad2/3 involved in Rnf111-mediated HSPC development. In conclusion, our study demonstrated that Rnf111 contributes to the development of HSPC by maintaining Smad2/3 phosphorylation and the Gcsfr/NO signaling pathway activation. Keywords: Rnf111, Ubiquitin ligase (UbL), HSPC, Smad2/3, Gcsfr/NO.

中文翻译：

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Rnf111 在斑马鱼中通过 Smad2/3-Gcsfr/NO 轴在调节确定性造血干细胞和祖细胞的发育中起关键作用。


造血相关因子的泛素化或 SUMO化在造血中起关键作用。RNF111 被称为泛素连接酶 （Ubl），是一种新发现的 SUMO 靶向泛素连接酶 （STUbl），参与 TGF-β 家族成员介导的多种信号通路。然而，它在造血中的作用仍不清楚。在此，通过 CRISPR/Cas9 介导的基因组编辑产生了可遗传的 Rnf111 突变斑马鱼系。在 Rnf111 缺陷突变体中发现确定性造血的造血干细胞和祖细胞 （HSPC） 受损。Rnf111 的消融导致 HSPC 中 Smad2/3 的磷酸化降低。确定性内胚层 2 诱导剂 （IDE2） 特异性激活 TGF-β 信号传导和下游 Smad2 磷酸化，可以恢复 Rnf111 缺陷胚胎的最终造血。进一步的分子机制研究显示，Gcsfr/NO 信号转导是 Smad2/3 参与 Rnf111 介导的 HSPC 发育的重要靶途径。总之，我们的研究表明，Rnf111 通过维持 Smad2/3 磷酸化和 Gcsfr/NO 信号通路激活促进 HSPC 的发展。关键字：Rnf111， 泛素连接酶 （UbL）， HSPC， Smad2/3， Gcsfr/NO.