Wnt/β-连环素信号通路是一个至关重要的保守途径，对于发育、稳态和肿瘤发生起着关键作用。其调节剂的发现在基础和转化研究中具有巨大价值。通过筛选，我们确定了脱泛素酶USP10作为β-连环素的一个关键调节剂。在机制上，USP10通过保守的结构域与重要的支架蛋白Axin1结合，并通过K48链的脱泛素化作用稳定Axin1。令人惊讶的是，USP10使Axin1和β-连环素发生物理连接，并促进β-连环素抑制的相分离，无论是否存在酶活性。在功能上，USP10的酶活性主要调节胚胎发育，并通过对抗β-连环素来共同调控肠道稳态。在结直肠癌中，USP10通过促进相分离显著抑制癌细胞生长，并在临床上与Wnt/β-连环素的强度相关。综上所述，我们发现USP10在多个生物过程中对抗β-连环素并揭示了酶依赖和非酶依赖的“双调节”机制。这两个USP10的功能并行工作并且与环境有关。版权所有 © 2023 Elsevier Ltd. 保留所有权利。

Wnt/β-catenin signaling is a conserved pathway crucially governing development, homeostasis, and oncogenesis. Discoveries of its regulators hold great values in both basic and translational research. Through screening, we identified a deubiquitinase, USP10, as a critical modulator of β-catenin. Mechanistically, USP10 binds to key scaffold Axin1 via conserved motifs and stabilizes Axin1 through K48-linked deubiquitination. Surprisingly, USP10 physically tethers Axin1 and β-catenin and promotes the phase separation for β-catenin suppression regardless of the enzymatic activity. Function-wise, USP10 enzymatic activity preferably regulates embryonic development and both the enzymatic activity and physical function jointly control intestinal homeostasis by antagonizing β-catenin. In colorectal cancer, USP10 substantially represses cancer growth mainly through physical promotion of phase separation and correlates with Wnt/β-catenin magnitude clinically. Collectively, we discovered USP10 functioning in multiple biological processes against β-catenin and unearthed the enzyme-dependent and -independent "dual-regulating" mechanism. These two functions of USP10 work in parallel and are context dependent.Copyright © 2023 Elsevier Ltd. All rights reserved.