Evidence for NL1-independent nuclear translocation of the mineralocorticoid receptor.
In the absence of hormone, corticosteroid receptors are primarily located in the cytoplasm, and they rapidly accumulate in the nucleus (t0.5 = 5 min) upon ligand binding. It is generally believed that the dissociation of hsp90 from the receptor is an absolute requirement for allowing its nuclear translocation. However, recent evidence suggests that hsp90 may remain associated with the glucocorticoid receptor during this process, and thus, the receptor nuclear localization signal (NLS) is not obscured by its presence. To determine the requirements for mineralocorticoid receptor (MR) nuclear transport, it was first shown that in rat kidney collecting duct cells, nuclear localization of MR in the presence of aldosterone was complete in 10 min. Although the hsp90 inhibitor radicicol delayed nuclear translocation, it did not prevent complete nuclear accumulation of MR at longer incubation times (t0.5 = 30-40 min). MR carbamylation generates a non-steroid-transformed receptor that, in contrast to native MR, is very stable in cell-free systems. In contrast to the full nuclear translocation of aldosterone-transformed MR, only a fraction of the carbamylated MR became nuclear in digitonin-permeabilized cells even though its NLS is exposed. Furthermore, while preincubation of permeabilized cells with NL1 peptide or anti-NL1 antibody fully inhibited the nuclear translocation of NL1-tagged albumin, neither treatment fully inhibited MR nuclear translocation. We postulate that there are at least two possible mechanisms for MR nuclear translocation. One of them is hsp90- and NL1-dependent, and the other functions in a manner that is independent of the classical pathway.
Piwien Pilipuk
Graciela
G
Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina.
Vinson
Gavin P
GP
Sanchez
Celso Gomez
CG
Galigniana
Mario D
MD
D021581:Active Transport, Cell Nucleus; D000450:Aldosterone; D000818:Animals; D018841:HSP90 Heat-Shock Proteins; D007668:Kidney / Q000166:cytology; D007700:Kinetics; D019913:Nuclear Localization Signals; D051381:Rats; D018161:Receptors, Mineralocorticoid / Q000378:metabolism*
D018841:HSP90 Heat-Shock Proteins; D019913:Nuclear Localization Signals; D018161:Receptors, Mineralocorticoid; D000450:Aldosterone