Abstract
Cell-based high-throughput screens (HTSs) targeting heterologously expressed proteins in yeast identify compounds that often display relevant biological activity when tested in cell culture. We developed a fission yeast-based HTS to detect small-molecule inhibitors of mammalian cyclic nucleotide phosphodiesterases (PDEs). These screens are carried out in Schizosaccharomyces pombe using a PKA-repressed fbp1-ura4 reporter whose expression due to low PKA activity prevents cells from growing in medium containing the pyrimidine analog 5-fluoro orotic acid (5FOA). We describe here the steps required to construct strains for screening and to optimize conditions for successful screens.
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Acknowledgments
This work was supported by NIH grant GM079662, the Peter Rieser Lectureship Fund, and a grant from Boston College to C.S.H.
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de Medeiros, A.S., Hoffman, C.S. (2015). A Yeast-Based High-Throughput Screen for Modulators of Phosphodiesterase Activity. In: Zaccolo, M. (eds) cAMP Signaling. Methods in Molecular Biology, vol 1294. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2537-7_14
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DOI: https://doi.org/10.1007/978-1-4939-2537-7_14
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