A yeast-based high-throughput screen for modulators of phosphodiesterase activity.

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.

DOI: 10.1007/978-1-4939-2537-7_14

Cite this paper

@article{Medeiros2015AYH, title={A yeast-based high-throughput screen for modulators of phosphodiesterase activity.}, author={Ana Santos de Medeiros and Charles S Hoffman}, journal={Methods in molecular biology}, year={2015}, volume={1294}, pages={181-90} }