Evidence that the peptidylprolyl isomerase domain of the hsp90-binding immunophilin FKBP52 is involved in both dynein interaction and glucocorticoid receptor movement to the nucleus.
We have previously shown that immunoadsorption of the FKBP52 immunophilin component of steroid receptor.hsp90 heterocomplexes is accompanied by coadsorption of cytoplasmic dynein, a motor protein involved in retrograde transport of vesicles toward the nucleus. Coimmunoadsorption of dynein is competed by an expressed fragment of FKBP52 comprising its peptidylprolyl isomerase (PPIase) domain (Silverstein, A. M., Galigniana, M. D., Kanelakis, K. C., Radanyi, C., Renoir, J.-M., and Pratt, W. B. (1999) J. Biol. Chem. 52, 36980-36986). Here we show that cotransfection of 3T3 cells with the FKBP52 PPIase domain and a green fluorescent protein (GFP) glucocorticoid receptor (GR) chimera inhibits dexamethasone-dependent movement of the GFP-GR from the cytoplasm to the nucleus. Cotransfection with FKBP12 does not affect GFP-GR movement. Inhibition of movement by the FKBP52 PPIase domain is abrogated in cells treated with colcemid to eliminate microtubules prior to steroid addition. After withdrawal of colcemid, microtubules reform, and PPIase inhibition of GFP-GR movement is restored. These observations are consistent with the notion that FKBP52 targets retrograde movement of the GFP-GR along microtubules by linking the receptor to the dynein motor. Here, we also show that native GR.hsp90 heterocomplexes immunoadsorbed from L cell cytosol contain dynein and that GR.hsp90 heterocomplexes assembled in reticulocyte lysate contain cytoplasmic dynein in a manner that is competed by the PPIase domain of FKBP52.
Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
M D
MD
Galigniana
C
C
Radanyi
J M
JM
Renoir
P R
PR
Housley
W B
WB
Pratt
D016475:3T3 Cells; D000818:Animals; D016227:Benzoquinones; D002467:Cell Nucleus / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D047029:Lactams, Macrocyclic; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D019696:Peptidylprolyl Isomerase / Q000378:metabolism*; D021381:Protein Transport; D011809:Quinones / Q000494:pharmacology; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D022021:Tacrolimus Binding Proteins / Q000737:chemistry / Q000378:metabolism*
D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D008164:Luminescent Proteins; D011809:Quinones; D011965:Receptors, Glucocorticoid; D049452:Green Fluorescent Proteins; D004398:Dyneins; D022021:Tacrolimus Binding Proteins; C403485:tacrolimus binding protein 4; D019696:Peptidylprolyl Isomerase; C001277:geldanamycin