title: Intracellular distribution of a cytoplasmic progesterone receptor mutant and of immunophilins cyclophilin 40 and FKBP59: effects of cyclosporin A, of various metabolic inhibitors and of several culture conditions.
abstract: The effect of cyclosporin A (CsA) on the intracellular distribution of a mutated NLS minus rabbit progesterone receptor (PRm) and the receptor-associated immunophilins, cyclophilin 40 (Cyp40) and FKBP59, was tested in Lc13 cells by indirect immunofluorescent staining. PRm, which is cytoplasmic in absence of progesterone, is shifted to the nucleus by the hormone as well as by CsA, but not by FK506 or Rapamycin [I. Jung-Testas, M.-C. Lebeau, E.E. Baulieu. C.R. Acad. Sci. Paris 318 (1995) 873-878]. However the time course of nuclear import due to CsA and its sensitivity to N-ethyl maleimide (NEM) and to a calmodulin inhibitor (W7) was different from those observed for the hormonal effect. Cyp40 in Lc13 cells is localized mainly in the nucleoli. CsA treatment increased nucleolar staining, while NEM and W7 caused it to decrease; after actinomycin D (1 microM) nucleolar staining of Cyp40 disappeared. FKBP59 is mainly cytoplasmic and concentrated in the perinuclear region, never in the nucleoli. CsA, actino D and W7 treatment did not influence FKBP59 localization. In serum-deprived medium FKBP59 was cytoplasmic, but when the culture medium was enriched (20% serum, insulin and EGF) FKBP59 became perinuclear and hsp 86 was partly shifted to the nucleus, but PRm remained cytoplasmic. CsA has an effect on PRm distribution, while it does not influence Cyp40 and FKBP59 localization. In presence of actino D the labelling of Cyp40 disappears from the nucleoli, while the distribution of PRm and FKBP59 is unaffected. Growth factors influence FKBP59 but not PRm or Cyp40. These results suggest that these proteins shuttle independently and that their association is transient.
authors.affiliation: INSERM U 33 and U 488, Hôpital de Bicêtre, Le Kremlin Bicêtre, France.
authors.forename: M C
authors.initials: MC
authors.lastname: Lebeau
authors.forename: I
authors.initials: I
authors.lastname: Jung-Testas
authors.forename: E E
authors.initials: EE
authors.lastname: Baulieu
mesh_terms: D000818:Animals; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002352:Carrier Proteins / Q000378:metabolism*; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins; D016572:Cyclosporine / Q000494:pharmacology*; D003593:Cytoplasm / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006358:Hot Temperature; D020104:Immunophilins / Q000378:metabolism*; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007739:L Cells; D051379:Mice; D016296:Mutagenesis; D019696:Peptidylprolyl Isomerase / Q000378:metabolism*; D010643:Phenoxybenzamine / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology*; D011817:Rabbits; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins; D017384:Sequence Deletion; D020123:Sirolimus / Q000494:pharmacology; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D022021:Tacrolimus Binding Proteins; D014162:Transfection
chemical_list: D002147:Calmodulin; D002352:Carrier Proteins; D000081406:Peptidyl-Prolyl Isomerase F; D007166:Immunosuppressive Agents; C000631446:PPIF protein, mouse; D011980:Receptors, Progesterone; D011994:Recombinant Proteins; D013449:Sulfonamides; D010643:Phenoxybenzamine; D003609:Dactinomycin; D011374:Progesterone; C017967:W 7; D016572:Cyclosporine; D021983:Cyclophilins; D022021:Tacrolimus Binding Proteins; D020104:Immunophilins; D019696:Peptidylprolyl Isomerase; D005033:Ethylmaleimide; D020123:Sirolimus