Abstract
Kinase cascades are a fundamental feature of cellular signaling and play a vital role in disease progression. Thus, tools to monitor the activity of kinase cascades are of high importance. Our group has developed a split-luciferase biosensor system to monitor the activity of the Hippo pathway, a kinase cascade that regulates a wide variety of cellular processes. We have applied this biosensor for high-throughput screening to discover novel upstream kinase regulators of Hippo signaling. In this chapter, we describe our approach to designing, validating, and using the biosensor for screening procedures, which provides an example for the reader should they wish to design a similar biosensor system for their own purposes.
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References
Badr CE, Tannous BA (2011) Bioluminescence imaging: progress and applications. Trends Biotechnol 29:624–633
Xu T, Close D, Handagama W et al (2016) The expanding toolbox of in vivo bioluminescent imaging. Front Oncol 6:150
Azad T, Tashakor A, Hosseinkhani S (2014) Split-luciferase complementary assay: applications, recent developments, and future perspectives. Anal Bioanal Chem 406:5541–5560
Hall MP, Unch J, Binkowski BF et al (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7:1848–1857
Dixon AS, Schwinn MK, Hall MP et al (2016) NanoLuc complementation reporter optimized for accurate measurement of protein interactions in cells. ACS Chem Biol 11:400–408
England CG, Ehlerding EB, Cai W (2016) NanoLuc: a small luciferase is brightening up the field of bioluminescence. Bioconjug Chem 27:1175–1187
Wu Z, Guan KL (2021) Hippo signaling in embryogenesis and development. Trends Biochem Sci 46:51–63
Dey A, Varelas X, Guan KL (2020) Targeting the hippo pathway in cancer, fibrosis, wound healing and regenerative medicine. Nat Rev Drug Discov 19:480–494
Cao X, Wang C, Liu J et al (2020) Regulation and functions of the hippo pathway in stemness and differentiation. Acta Biochim Biophys Sin 52:736–748
Pan D (2010) The hippo signaling pathway in development and cancer. Dev Cell 19:491–505
Ma S, Meng Z, Chen R et al (2019) The hippo pathway: biology and pathophysiology. Annu Rev Biochem 88:577–604
Dong J, Feldmann G, Huang J et al (2007) Elucidation of a universal size-control mechanism in drosophila and mammals. Cell 130:1120–1133
Sanchez-Vega F, Mina M, Armenia J et al (2018) Oncogenic signaling pathways in the cancer genome atlas. Cell 173:321–337.e10
Xu T, Wang W, Zhang S et al (1995) Identifying tumor suppressors in genetic mosaics: the drosophila lats gene encodes a putative protein kinase. Development 121:1053–1063
St John MAR, Tao W, Fei X et al (1999) Mice deficient of Lats1 develop soft-tissue sarcomas, ovarian tumours and pituitary dysfunction. Nat Genet 21:182–186
Nouri K, Azad T, Lightbody E et al (2019) A kinome-wide screen using a NanoLuc LATS luminescent biosensor identifies ALK as a novel regulator of the hippo pathway in tumorigenesis and immune evasion. FASEB J 33:12487–12499
Nouri K, Azad T, Ling M et al (2019) Identification of celastrol as a novel YAP-TEAD inhibitor for cancer therapy by high throughput screening with ultrasensitive YAP/TAZ–TEAD biosensors. Cancers (Basel) 11:1596
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Pipchuk, A., Yang, X. (2022). Monitoring Hippo Signaling Pathway Activity Using a Luciferase-Based Large Tumor Suppressor (LATS) Biosensor. In: Kim, SB. (eds) Bioluminescence. Methods in Molecular Biology, vol 2525. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2473-9_11
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DOI: https://doi.org/10.1007/978-1-0716-2473-9_11
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