Metazoan development relies on intricate cell differentiation, communication, and migration pathways, which ensure proper formation of specialized cell types, tissues, and organs. These pathways are crucially controlled by ubiquitylation, a reversible post-translational modification that regulates the stability, activity, localization, or interaction landscape of substrate proteins. Specificity of ubiquitylation is ensured by E3 ligases, which bind substrates and co-operate with E1 and E2 enzymes to mediate ubiquitin transfer. Cullin3-RING ligases (CRL3s) are a large class of multi-subunit E3s that have emerged as important regulators of cell differentiation and development. In particular, recent evidence from human disease genetics, animal models, and mechanistic studies have established their involvement in the control of craniofacial and brain development. Here, we summarize regulatory principles of CRL3 assembly, substrate recruitment, and ubiquitylation that allow this class of E3s to fulfill their manifold functions in development. We further review our current mechanistic understanding of how specific CRL3 complexes orchestrate neuroectodermal differentiation and highlight diseases associated with their dysregulation. Based on evidence from human disease genetics, we propose that other unknown CRL3 complexes must help coordinate craniofacial and brain development and discuss how combining emerging strategies from the field of disease gene discovery with biochemical and human pluripotent stem cell approaches will likely facilitate their identification.

后生动物的发育依赖于复杂的细胞分化、通讯和迁移途径，确保特殊细胞类型、组织和器官的正确形成。这些途径主要受到泛素化的控制，泛素化是一种可逆的翻译后修饰，可调节底物蛋白的稳定性、活性、定位或相互作用。E3 连接酶确保泛素化的特异性，E3 连接酶结合底物并与 E1 和 E2 酶配合介导泛素转移。Cullin3-RING 连接酶 (CRL3) 是一大类多亚基 E3，已成为细胞分化和发育的重要调节因子。特别是，最近来自人类疾病遗传学、动物模型和机制研究的证据已经证实它们参与颅面和大脑发育的控制。在这里，我们总结了 CRL3 组装、底物招募和泛素化的调控原理，这些原理使此类 E3 在发育中能够实现其多种功能。我们进一步回顾了目前对特定 CRL3 复合物如何协调神经外胚层分化的机制理解，并强调了与其失调相关的疾病。基于人类疾病遗传学的证据，我们提出其他未知的 CRL3 复合物必须有助于协调颅面和大脑发育，并讨论如何将疾病基因发现领域的新兴策略与生化和人类多能干细胞方法相结合，以促进其识别。