title: Screening for a gene deletion mutant whose temperature sensitivity is suppressed by FK506 in budding yeast and its application for a positive screening for drugs inhibiting calcineurin.
abstract: Calcineurin, which is a Ca(2+)/calmodulin-dependent protein phosphatase, is a key mediator in calcium signaling in diverse biological processes and of clinical importance as the target of the immunosuppressant FK506. To identify a mutant(s) in which calcineurin is activated, inhibiting cellular growth as a result, we screened for a mutant(s) whose temperature sensitivity would be suppressed by FK506 from the budding yeast non-essential gene deletion library. We found that the temperature sensitivity of cells in which the conserved Verprolin VRP1 gene had been deleted, which gene is required for actin organization and endocytosis, was suppressed by either FK506 or by cnb1 deletion. Indeed, the calcineurin activity increased significantly in the ∆vrp1 cells. Finally, we demonstrated that the ∆vrp1 strain to be useful as an indicator in a positive screening for bioactive compounds inhibiting calcineurin.
authors.affiliation: a Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter , Hiroshima University , 1-3-1 kagamiyama, Higashi-Hiroshima , 739-8530 Japan.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Kume
authors.forename: Takayuki
authors.initials: T
authors.lastname: Koyano
authors.forename: Junpei
authors.initials: J
authors.lastname: Takata
authors.forename: Ko
authors.initials: K
authors.lastname: Wakabayashi
authors.forename: Masaki
authors.initials: M
authors.lastname: Mizunuma
authors.forename: Tokichi
authors.initials: T
authors.lastname: Miyakawa
authors.forename: Dai
authors.initials: D
authors.lastname: Hirata
mesh_terms: D019703:Calcineurin / Q000235:genetics / Q000378:metabolism; D065095:Calcineurin Inhibitors / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D017353:Gene Deletion; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology*; D013696:Temperature
chemical_list: D065095:Calcineurin Inhibitors; D008840:Microfilament Proteins; D029701:Saccharomyces cerevisiae Proteins; C090350:VRP1 protein, S cerevisiae; D019703:Calcineurin; D016559:Tacrolimus
keywords: FK506; calcineurin; drug screening; yeast