State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 5626 Renmin Street, Changchun, Jilin 130022, P.R. China.
Nanoscale. 2017 Nov 9;9(43):16993-17003. doi: 10.1039/c7nr05818g.
Yes-associated protein (YAP) is well known for being an effecter of the Hippo signaling cascade that plays a critical role in organ size control, tumorigenesis, and regeneration. As YAP is a transcriptional coactivator, nuclear accumulation is a crucial determinant of its function. Numerous investigations have provided insights into the regulation of YAP, such as upstream molecules of the Hippo pathway, cell contact inhibition, and mechanical forces. However, detailed information regarding YAP spatial localization and organization in cells remains uncertain, and how mechanical signals control YAP distribution and function is not fully known. Therefore, we used one of the super-resolution imaging techniques, direct stochastic optical reconstruction microscopy (dSTORM), combined with confocal microscopy, to solve these problems. We found that YAP is mainly distributed in clusters in the cells, and that both cell contact and pressure on cell surfaces promote the nuclear-to-cytoplasm translocation of YAP and its phosphorylation, but weaken the clustering of nuclear YAP and its transcriptional activity. Moreover, we found that pressure regulation may be more effective on YAP from cancer cells as compared to normal cells, which could help open a door to target YAP for anticancer drug design.
Yes 相关蛋白 (YAP) 作为 Hippo 信号级联反应的效应因子而广为人知，该级联反应在器官大小控制、肿瘤发生和再生中起着关键作用。由于 YAP 是一种转录共激活因子，核积累是其功能的关键决定因素。许多研究为 YAP 的调节提供了深入的了解，例如 Hippo 途径的上游分子、细胞接触抑制和机械力。然而，关于 YAP 在细胞中的空间定位和组织的详细信息仍然不确定，并且机械信号如何控制 YAP 的分布和功能也不完全清楚。因此，我们使用超分辨率成像技术之一，直接随机光学重建显微镜 (dSTORM)，结合共聚焦显微镜，来解决这些问题。我们发现 YAP 主要分布在细胞中的簇中，细胞接触和细胞表面的压力促进 YAP 的核质易位及其磷酸化，但削弱核 YAP 的聚类及其转录活性。此外，我们发现与正常细胞相比，压力调节对癌细胞中的 YAP 可能更有效，这可能有助于为抗癌药物设计靶向 YAP 打开一扇门。
