title: Fission yeast-based high-throughput screens for PKA pathway inhibitors and activators.
abstract: Features of the fission yeast Schizosaccharomyces pombe cAMP/PKA pathway make S. pombe particularly amenable for heterologous expression of cAMP pathway proteins such as GαS subunits and their cognate adenylyl cyclases, PKA catalytic and regulatory subunits, and cyclic nucleotide phosphodiesterases. We have constructed two PKA-repressed reporters for use in high-throughput screens to detect compounds that elevate or reduce PKA activity, thus facilitating the discovery of both inhibitors and activators of these target proteins. Here, we describe steps to construct screening strains and to optimize and conduct these screens.
authors.affiliation: Biology Department, Boston College, 140 Commonwealth Avenue, Higgins Hall 401b, Chestnut Hill, MA, 02467, USA.
authors.forename: Ana Santos
authors.initials: AS
authors.lastname: de Medeiros
authors.forename: Grace
authors.initials: G
authors.lastname: Kwak
authors.forename: Jordan
authors.initials: J
authors.lastname: Vanderhooft
authors.forename: Sam
authors.initials: S
authors.lastname: Rivera
authors.forename: Rachel
authors.initials: R
authors.lastname: Gottlieb
authors.forename: Charles S
authors.initials: CS
authors.lastname: Hoffman
mesh_terms: D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D057166:High-Throughput Screening Assays; D047908:Intracellular Signaling Peptides and Proteins / Q000494:pharmacology*; D012568:Schizosaccharomyces / Q000187:drug effects* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C015482:protein kinase modulator; D017868:Cyclic AMP-Dependent Protein Kinases