Abstract
The Hippo signaling network is dependent on protein–protein interactions (PPIs) as a mechanism of signal transduction to regulate organ size, cellular proliferation and differentiation, tumorigenesis, and other cellular processes. Current efforts aim to resolve the complex regulation of upstream Hippo components or focus on identifying targeted drugs for use in cancer therapy. Despite extensive characterization of the Hippo pathway interactome by affinity purification mass spectrometry (AP-MS) and other methodologies, previous research methods have not been sufficient to achieve these aims. In this review, we describe several recent studies that make use of luciferase-based biosensors as a new approach to study the Hippo Pathway. These biosensors serve as powerful tools with which to study PPIs both in vitro using purified biosensor proteins, and in real time in live cells. Notably, luciferase biosensors have excellent sensitivity and have been used to screen for upstream kinase regulators of the Hippo pathway. Furthermore, the high sensitivity and stability of these biosensors enables their application in high throughput screening for Hippo-targeted chemotherapeutics. Finally, we describe the strengths and weaknesses of this method relative to AP-MS and discuss potential future directions for using biosensors to study Hippo signaling.
Highlights
The Hippo PathwayThe Hippo pathway is an evolutionarily conserved signaling cascade that plays central roles in human physiology and disease (Pan, 2010; Ma et al, 2019)
Phosphorylated YAP and TAZ (YAP/TAZ) is sequestered in the cytoplasm and is unable to associate with the TEAD family of transcription factors in the nucleus to direct transcription of Hippo-associated genes
In 2018, we presented research that made use of a firefly luciferase (FLuc)based LATS biosensor to identify VEGFR as an upstream regulator of Hippo signaling (Azad et al, 2018)
Summary
Using Biosensors to Study Protein–Protein Interaction in the Hippo Pathway. We describe several recent studies that make use of luciferasebased biosensors as a new approach to study the Hippo Pathway. These biosensors serve as powerful tools with which to study PPIs both in vitro using purified biosensor proteins, and in real time in live cells. Luciferase biosensors have excellent sensitivity and have been used to screen for upstream kinase regulators of the Hippo pathway. We describe the strengths and weaknesses of this method relative to AP-MS and discuss potential future directions for using biosensors to study Hippo signaling
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