Toh Pearlyn Jia Ying
Mechanobiology Institute, National University of Singapore, Singapore, Singapore.
Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2025;2840:75-87. doi: 10.1007/978-1-0716-4047-0_6.
YAP is a central regulator of the Hippo-YAP signaling axis, an evolutionarily conserved pathway that modulates organ growth and regeneration. Dysregulation of YAP signaling leads to uncontrolled proliferation, promoting epithelial-to-mesenchymal transition and invasion in cancer metastasis. Exogenous manipulation of YAP activity at the second-to-minute timescale is an important step in studying the signaling pathway. We present an optogenetic system to control the subcellular localization of YAP and therefore its activity as a transcriptional co-activator. We used the LOV2-Jα interacting domains to photocage a nuclear localization signal (NLS) attached to YAP. Under 488 nm light, the Jα helix unfolds and the interaction with LOV2 is disrupted, thereby exposing the NLS and allowing for the entire optogenetic construct to be shuttled into the nucleus. This nuclear translocation is reversible and tuneable and demonstrates functional activity after nuclear localization both in vitro and in vivo.
YAP是Hippo-YAP信号轴的核心调节因子，这是一条在进化上保守的途径，可调节器官生长和再生。YAP信号失调会导致不受控制的增殖，促进上皮-间质转化和癌症转移中的侵袭。在秒到分钟的时间尺度上对外源YAP活性进行操作是研究该信号通路的重要一步。我们提出了一种光遗传学系统，以控制YAP的亚细胞定位，从而控制其作为转录共激活因子的活性。我们使用LOV2-Jα相互作用结构域对附着于YAP的核定位信号（NLS）进行光笼化。在488nm光下，Jα螺旋展开，与LOV2的相互作用被破坏，从而暴露NLS，并使整个光遗传学构建体穿梭进入细胞核。这种核转位是可逆的且可调节的，并且在体外和体内核定位后均表现出功能活性。
