title: Rapamycin-binding FKBP25 associates with diverse proteins that form large intracellular entities.
abstract: In this paper, we show some evidence that a member of the FK506-binding proteins, FKBP25 is associated to diverse components that are part of several different intracellular large-molecular mass entities. The FKBP25 is a high-affinity rapamycin-binding immunophilin, which has nuclear translocation signals present in its PPIase domain but it was detected both in the cytoplasm compartment and in the nuclear proteome. Analyses of antiFKBP25-immunoprecipitated proteins have revealed that the endogenous FKBP25 is associated to the core histones of the nucleosome, and with several proteins forming spliceosomal complexes and ribosomal subunits. Using polyclonal antiFKBP25 we have detected FKBP25 associated with polyribosomes. Added RNAs or 0.5M NaCl release FKBP25 that was associated with the polyribosomes indicating that the immunophilin has an intrinsic capacity to form complexes with polyribonucleotides via its charged surface patches. Rapamycin or FK506 treatments of the polyribosomes isolated from porcine brain, HeLa and K568 cells caused a residual release of the endogenous FKBP25, which suggests that the immunophilin also binds to some proteins via its PPIase cavity. Our proteomics study indicates that the nuclear pool of the FKBP25 targets various nuclear proteins that are crucial for packaging of DNA, chromatin remodeling and pre-mRNA splicing whereas the cytosolic pool of this immunophilin is bound to some components of the ribosome.
authors.affiliation: Commissariat à l'Energie Atomique, Direction des Sciences du Vivant, Institut de Biologie et de Technologies de Saclay, Service d'Ingénierie Moléculaire des Protéines, Gif sur Yvette, France. Electronic address: galat@dsvidf.cea.fr.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Galat
authors.affiliation: Commissariat à l'Energie Atomique, Direction des Sciences du Vivant, Institut de Biologie et de Technologies de Saclay, Service d'Ingénierie Moléculaire des Protéines, Gif sur Yvette, France.
authors.forename: Robert
authors.initials: R
authors.lastname: Thai
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D011485:Protein Binding; D012270:Ribosomes / Q000378:metabolism; D013552:Swine; D022021:Tacrolimus Binding Proteins / Q000378:metabolism*
chemical_list: C074856:FKBP3 protein, human; D022021:Tacrolimus Binding Proteins
keywords: FKBP25; FKBPs; Immunophilin; PPIase; Rapamycin