Leveraging the extraordinary potential of human pluripotent stem cells (hPSCs) requires an understanding of the mechanisms underlying cell-fate decisions. Substrate elasticity can induce differentiation by signaling through the transcriptional coactivator Yes-associated protein (YAP). Cells cultured on surfaces mimicking brain elasticity exclude YAP from their nuclei and differentiate to neurons. How YAP localization is controlled during neural differentiation has been unclear. We employed CRISPR/Cas9 to tag endogenous YAP in hPSCs and used this fusion protein to identify YAP's interaction partners. This engineered cell line revealed that neural differentiation promotes a change in YAP interactors, including a dramatic increase in angiomotin (AMOT) interaction with YAP. AMOT regulates YAP localization during differentiation. AMOT expression increases during neural differentiation and leads to YAP nuclear exclusion. Our findings that AMOT-dependent regulation of YAP helps direct hPSC fate provide insight into the molecular mechanisms by which the microenvironment can induce neural differentiation.

利用人类多能干细胞（hPSC）的巨大潜力，需要了解细胞命运决定的潜在机制。底物弹性可通过转录共激活因子Yes相关蛋白（YAP）发出信号来诱导分化。在模仿大脑弹性的表面上培养的细胞将YAP从其细胞核中排除，并分化为神经元。尚不清楚神经分化过程中如何控制YAP定位。我们使用CRISPR / Cas9标记hPSC中的内源性YAP，并使用该融合蛋白鉴定YAP的相互作用伙伴。这种工程改造的细胞系揭示了神经分化促进了YAP相互作用因子的变化，包括与YAP的血管动蛋白（AMOT）相互作用的急剧增加。AMOT在分化过程中调节YAP的定位。AMOT表达在神经分化过程中增加，并导致YAP核排斥。我们的研究发现，依赖AMOT的YAP调节有助于指导hPSC的命运，从而提供了对微环境可以诱导神经分化的分子机制的深刻见解。