title: FK506 binding protein 12 mediates sensitivity to both FK506 and rapamycin in murine mast cells.
abstract: The immunosuppressive drugs FK506 and rapamycin bind to a family of intracellular proteins termed FK506-binding proteins (FKBP). FK506 and rapamycin inhibit lymphocyte-activation pathways by forming complexes with an FKBP; subsequently, the drug/FKBP complexes interact with target molecules involved in signal transduction. A key target of FK506/FKBP12 complexes is calcineurin, a calcium- and calmodulin-dependent serine/threonine phosphatase. In mammalian cells, rapamycin treatment is associated with inhibition of the activity of several cellular serine/threonine kinases, including p70 S6 kinase. These kinases may function in signaling pathways involving TOR gene producs, which have been shown to interact with rapamycin/FKBP12 complexes in vitro. To determine if FKBP12 mediates the effects of both FK506 and rapamycin in mammalian cells, we overexpressed FKBP12 in a murine mast cell line. Increased expression of FKBP12 resulted in increased sensitivity to FK506 and rapamycin, as measured by inhibition of calcineurin activity and p70 S6 kinase activity, respectively. In contrast, overexpression of FKBP25 had no effect on sensitivity to either drug. Two distinct point mutations in FKBP12, one altering a hydrophobic residue within the drug-binding pocket and the other changing a charged surface residue of FKBP12, abrogated its ability to mediate sensitivity to FK506 and rapamycin. These results establish that FKBP12 can mediate sensitivity to both FK506 and rapamycin in mammalian cells.
authors.affiliation: Division of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA 02115.
authors.forename: D A
authors.initials: DA
authors.lastname: Fruman
authors.forename: M A
authors.initials: MA
authors.lastname: Wood
authors.forename: C K
authors.initials: CK
authors.lastname: Gjertson
authors.forename: H R
authors.initials: HR
authors.lastname: Katz
authors.forename: S J
authors.initials: SJ
authors.lastname: Burakoff
authors.forename: B E
authors.initials: BE
authors.lastname: Bierer
mesh_terms: D000818:Animals; D001483:Base Sequence; D019703:Calcineurin; D002148:Calmodulin-Binding Proteins / Q000037:antagonists & inhibitors; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D004351:Drug Resistance; D006360:Heat-Shock Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008407:Mast Cells / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011090:Polyenes / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D019893:Ribosomal Protein S6 Kinases; D020123:Sirolimus; D016559:Tacrolimus / Q000494:pharmacology*; D022021:Tacrolimus Binding Proteins; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C489567:Fkbp3 protein, mouse; D006360:Heat-Shock Proteins; D011090:Polyenes; C074856:FKBP3 protein, human; D017346:Protein Serine-Threonine Kinases; D019893:Ribosomal Protein S6 Kinases; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D022021:Tacrolimus Binding Proteins; D020123:Sirolimus; D016559:Tacrolimus