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Glucocorticoid receptor (GR) is cytoplasmic in the absence of ligand and localizes to the nucleus after steroid binding. Previous evidence demonstrated that the hsp90-based heterocomplex bound to GR is required for the efficient retrotransport of the receptor to the nuclear compartment. We examined the putative association of GR and its associated chaperone heterocomplex with structures of the nuclear pore. We found that importin β and the integral nuclear pore glycoprotein Nup62 interact with hsp90, hsp70, p23, and the TPR domain proteins FKBP52 and PP5. Nup62 and GR were able to interact in a more efficient manner when chaperoned by the hsp90-based heterocomplex. Interestingly, the binding of hsp70 and p23 to Nup62 does not require the presence of hsp90, whereas the association of FKBP52 and PP5 is hsp90 dependent, as indicated by the results of experiments where the hsp90 function was disrupted with radicicol. The ability of both FKBP52 and PP5 to interact with Nup62 was abrogated in cells overexpressing the TPR peptide. Importantly, GR cross-linked to the hsp90 heterocomplex was able to translocate to the nucleus in digitonin-permeabilized cells treated with steroid, suggesting that GR could pass through the pore in its untransformed state. Copyright © 2009, American Society for Microbiology. All Rights Reserved.


Documento: Artículo
Título:Nuclear import of the glucocorticoid receptor-hsp90 complex through the nuclear pore complex is mediated by its interaction with Nup62 and importin β
Autor:Echeverría, P.C.; Mazaira, G.; Erlejman, A.; Gomez-Sanchez, C.; Pilipuk, G.P.; Galigniana, M.D.
Filiación:Fundación Instituto Leloir-IIBBA/CONICET, Buenos Aires, Argentina
Departamento de Química Biológica, Facultad de Ciencias Exactas Y Naturales, Universidad de Buenos Aires, Argentina
Division of Endocrinology, G. V. (Sonny) Montgomery VA Medical Center, University of Mississippi Medical Center, Jackson, MS, United States
Av. Patricias Argentinas 435, Buenos Aires (C1405BWE), Argentina
Palabras clave:digitonin; glucocorticoid receptor; glycoprotein; glycoprotein nup62; heat shock protein 70; heat shock protein 90; karyopherin beta; protein Fkbp52; protein p23; protein pp5; radicicol; steroid; tetratricopeptide repeat protein; unclassified drug; animal cell; article; cell membrane permeability; controlled study; human; human cell; mouse; nonhuman; nuclear import; nuclear pore; nuclear pore complex; priority journal; protein binding; protein cross linking; protein expression; protein function; protein protein interaction; structure analysis; tetratricopeptide repeat; Active Transport, Cell Nucleus; Animals; beta Karyopherins; Cell Line; Glycoproteins; HSP70 Heat-Shock Proteins; HSP90 Heat-Shock Proteins; Humans; Intramolecular Oxidoreductases; Mice; Multiprotein Complexes; Nuclear Pore; Nuclear Pore Complex Proteins; Receptors, Glucocorticoid; Tacrolimus Binding Proteins
Página de inicio:4788
Página de fin:4797
Título revista:Molecular and Cellular Biology
Título revista abreviado:Mol. Cell. Biol.
CAS:digitonin, 11024-24-1; radicicol, 12772-57-5; Glycoproteins; HSP70 Heat-Shock Proteins; HSP90 Heat-Shock Proteins; Intramolecular Oxidoreductases, 5.3.-; Multiprotein Complexes; NUP62 protein, mouse; Nuclear Pore Complex Proteins; Receptors, Glucocorticoid; Tacrolimus Binding Proteins, 5.2.1.-; beta Karyopherins; prostaglandin-E synthase,; tacrolimus binding protein 4, 5.2.1.-; tissue-factor-pathway inhibitor 2


  • Abbas-Terki, T., Donze, O., Briand, P.A., Picard, D., Hsp104 interacts with Hsp90 cochaperones in respiring yeast (2001) Mol. Cell. Biol, 21, pp. 7569-7575
  • Adam, S.A., Marr, R.S., Gerace, L., Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors (1990) J. Cell Biol, 111, pp. 807-816
  • Albermann, L., Shahin, V., Ludwig, Y., Schafer, C., Schillers, H., Oberleithner, H., Evidence for importin alpha independent nuclear translocation of glucocorticoid receptors in Xenopus laevis oocytes (2004) Cell. Physiol. Biochem, 14, pp. 343-350
  • Banerjee, A., Periyasamy, S., Wolf, I.M., Hinds Jr., T.D., Yong, W., Shou, W., Sanchez, E.R., Control of glucocorticoid and progesterone receptor subcellular localization by the ligand-binding domain is mediated by distinct interactions with tetratricopeptide repeat proteins (2008) Biochemistry, 47, pp. 10471-10480
  • Barsky, A., Gardy, J.L., Hancock, R.E., Munzner, T., Cerebral: A Cytoscape plugin for layout of and interaction with biological networks using subcellular localization annotation (2007) Bioinformatics, 23, pp. 1040-1042
  • Bukau, B., Horwich, A.L., The Hsp70 and Hsp60 chaperone machines (1998) Cell, 92, pp. 351-366
  • Cerami, E.G., Bader, G.D., Gross, B.E., Sander, C., cPath: Open source software for collecting, storing, and querying biological pathways (2006) BMC Bioinformatics, 7, p. 497
  • Conti, E., Izaurralde, E., Nucleocytoplasmic transport enters the atomic age (2001) Curr. Opin. Cell Biol, 13, pp. 310-319
  • Czar, M.J., Galigniana, M.D., Silverstein, A.M., Pratt, W.B., Geldanamycin, a heat shock protein 90-binding benzoquinone ansamycin, inhibits steroid-dependent translocation of the glucocorticoid receptor from the cytoplasm to the nucleus (1997) Biochemistry, 36, pp. 7776-7785
  • Davies, T.H., Ning, Y.M., Sanchez, E.R., A new first step in activation of steroid receptors: Hormone-induced switching of FKBP51 and FKBP52 immunophilins (2002) J. Biol. Chem, 277, pp. 4597-4600
  • Davies, T.H., Sanchez, E.R., Fkbp52 (2005) Int. J. Biochem. Cell Biol, 37, pp. 42-47
  • Denning, D.P., Patel, S.S., Uversky, V., Fink, A.L., Rexach, M., Disorder in the nuclear pore complex: The FG repeat regions of nucleoporins are natively unfolded (2003) Proc. Natl. Acad. Sci. USA, 100, pp. 2450-2455
  • Echeverria, P.C., Matrajt, M., Harb, O.S., Zappia, M.P., Costas, M.A., Roos, D.S., Dubremetz, J.F., Angel, S.O., Toxoplasma gondii Hsp90 is a potential drug target whose expression and subcellular localization are developmentally regulated (2005) J. Mol. Biol, 350, pp. 723-734
  • Freedman, N.D., Yamamoto, K.R., Importin 7 and importin alpha/importin beta are nuclear import receptors for the glucocorticoid receptor (2004) Mol. Biol. Cell, 15, pp. 2276-2286
  • Galigniana, M.D., Harrell, J.M., Housley, P.R., Patterson, C., Fisher, S.K., Pratt, W.B., Retrograde transport of the glucocorticoid receptor in neurites requires dynamic assembly of complexes with the protein chaperone hsp90 and is linked to the CHIP component of the machinery for proteasomal degradation (2004) Brain Res. Mol. Brain Res, 123, pp. 27-36
  • Galigniana, M.D., Harrell, J.M., Murphy, P.J., Chinkers, M., Radanyi, C., Renoir, J.M., Zhang, M., Pratt, W.B., Binding of hsp90-associated immunophilins to cytoplasmic dynein: Direct binding and in vivo evidence that the peptidylprolyl isomerase domain is a dynein interaction domain (2002) Biochemistry, 41, pp. 13602-13610
  • Galigniana, M.D., Harrell, J.M., O'Hagen, H.M., Ljungman, M., Pratt, W.B., Hsp90-binding immunophilins link p53 to dynein during p53 transport to the nucleus (2004) J. Biol. Chem, 279, pp. 22483-22489
  • Galigniana, M.D., Housley, P.R., DeFranco, D.B., Pratt, W.B., Inhibition of glucocorticoid receptor nucleocytoplasmic shuttling by okadaic acid requires intact cytoskeleton (1999) J. Biol. Chem, 274, pp. 16222-16227
  • Galigniana, M.D., Morishima, Y., Gallay, P.A., Pratt, W.B., Cyclophilin-A is bound through its peptidylprolyl isomerase domain to the cytoplasmic dynein motor protein complex (2004) J. Biol. Chem, 279, pp. 55754-55759
  • Galigniana, M.D., Radanyi, C., Renoir, J.M., Housley, P.R., Pratt, W.B., 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 (2001) J. Biol. Chem, 276, pp. 14884-14889
  • Gallo, L.I., Ghini, A.A., Pilipuk, G.P., Galigniana, M.D., Differential recruitment of tetratricopeptide repeat domain immunophilins to the mineralocorticoid receptor influences both heat-shock protein 90-dependent retrotransport and hormone-dependent transcriptional activity (2007) Biochemistry, 46, pp. 14044-14057
  • Georget, V., Terouanne, B., Nicolas, J.C., Sultan, C., Mechanism of antiandrogen action: Key role of hsp90 in conformational change and transcriptional activity of the androgen receptor (2002) Biochemistry, 41, pp. 11824-11831
  • Gilchrist, D., Mykytka, B., Rexach, M., Accelerating the rate of disassembly of karyopherin-cargo complexes (2002) J. Biol. Chem, 277, pp. 18161-18172
  • Grandi, P., Dang, T., Pane, N., Shevchenko, A., Mann, M., Forbes, D., Hurt, E., Nup93, a vertebrate homologue of yeast Nic96p, forms a complex with a novel 205-kDa protein and is required for correct nuclear pore assembly (1997) Mol. Biol. Cell, 8, pp. 2017-2038
  • Harrell, J.M., Kurek, I., Breiman, A., Radanyi, C., Renoir, J.M., Pratt, W.B., Galigniana, M.D., All of the protein interactions that link steroid receptor-hsp90-immunophilin heterocomplexes to cytoplasmic dynein are common to plant and animal cells (2002) Biochemistry, 41, pp. 5581-5587
  • Harrell, J.M., Murphy, P.J., Morishima, Y., Chen, H., Mansfield, J.F., Galigniana, M.D., Pratt, W.B., Evidence for glucocorticoid receptor transport on microtubules by dynein (2004) J. Biol. Chem, 279, pp. 54647-54654
  • Heitzer, M.D., Wolf, I.M., Sanchez, E.R., Witchel, S.F., DeFranco, D.B., Glucocorticoid receptor physiology (2007) Rev. Endocr. Metab. Disord, 8, pp. 321-330
  • Huang, T.W., Tien, A.C., Huang, W.S., Lee, Y.C., Peng, C.L., Tseng, H.H., Kao, C.Y., Huang, C.Y., POINT: A database for the prediction of protein-protein interactions based on the orthologous interactome (2004) Bioinformatics, 20, pp. 3273-3276
  • Jakel, S., Mingot, J.M., Schwarzmaier, P., Hartmann, E., Gorlich, D., Importins fulfill a dual function as nuclear import receptors and cytoplasmic chaperones for exposed basic domains (2002) EMBO J, 21, pp. 377-386
  • Kang, K.I., Devin, J., Cadepond, F., Jibard, N., Guiochon-Mantel, A., Baulieu, E.E., Catelli, M.G., In vivo functional protein-protein interaction: Nuclear targeted hsp90 shifts cytoplasmic steroid receptor mutants into the nucleus (1994) Proc. Natl. Acad. Sci. USA, 91, pp. 340-344
  • Madan, A.P., DeFranco, D.B., Bidirectional transport of glucocorticoid receptors across the nuclear envelope (1993) Proc. Natl. Acad. Sci. USA, 90, pp. 3588-3592
  • Mattaj, I.W., Englmeier, L., Nucleocytoplasmic transport: The soluble phase (1998) Annu. Rev. Biochem, 67, pp. 265-306
  • Moffatt, N.S., Bruinsma, E., Uhl, C., Obermann, W.M., Toft, D., Role of the cochaperone Tpr2 in Hsp90 chaperoning (2008) Biochemistry, 47, pp. 8203-8213
  • Murphy, P.J., Morishima, Y., Chen, H., Galigniana, M.D., Mansfield, J.F., Simons Jr., S.S., Pratt, W.B., Visualization and mechanism of assembly of a glucocorticoid receptor-Hsp70 complex that is primed for subsequent Hsp90-dependent opening of the steroid binding cleft (2003) J. Biol. Chem, 278, pp. 34764-34773
  • Nigg, E.A., Nucleocytoplasmic transport: Signals, mechanisms and regulation (1997) Nature, 386, pp. 779-787
  • Passinen, S., Valkila, J., Manninen, T., Syvala, H., Ylikomi, T., The C-terminal half of Hsp90 is responsible for its cytoplasmic localization (2001) Eur. J. Biochem, 268, pp. 5337-5342
  • Patel, S.S., Belmont, B.J., Sante, J.M., Rexach, M.F., Natively unfolded nucleoporins gate protein diffusion across the nuclear pore complex (2007) Cell, 129, pp. 83-96
  • Pemberton, L.F., Rosenblum, J.S., Blobel, G., Nuclear import of the TATA-binding protein: Mediation by the karyopherin Kap114p and a possible mechanism for intranuclear targeting (1999) J. Cell Biol, 145, pp. 1407-1417
  • Picard, D., Yamamoto, K.R., Two signals mediate hormonedependent nuclear localization of the glucocorticoid receptor (1987) EMBO J, 6, pp. 3333-3340
  • Pilipuk, G.P., Vinson, G.P., Sanchez, C.G., Galigniana, M.D., Evidence for NL1-independent nuclear translocation of the mineralocorticoid receptor (2007) Biochemistry, 46, pp. 1389-1397
  • Piwien-Pilipuk, G., Galigniana, M.D., Oxidative stress induced by L-buthionine-(S,R)-sulfoximine, a selective inhibitor of glutathione metabolism, abrogates mouse kidney mineralocorticoid receptor function (2000) Biochim. Biophys. Acta, 1495, pp. 263-280
  • Piwien-Pilipuk, G., Kanelakis, K.C., Ghini, A.A., Lantos, C.P., Litwack, G., Burton, G., Galigniana, M.D., Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone (2002) Biochim. Biophys. Acta, 1589, pp. 31-48
  • Pratt, W.B., Galigniana, M.D., Morishima, Y., Murphy, P.J., Role of molecular chaperones in steroid receptor action (2004) Essays Biochem, 40, pp. 41-58
  • Pratt, W.B., Krishna, P., Olsen, L.J., Hsp90-binding immunophilins in plants: The protein movers (2001) Trends Plant Sci, 6, pp. 54-58
  • Savory, J.G., Hsu, B., Laquian, I.R., Giffin, W., Reich, T., Hache, R.J., Lefebvre, Y.A., Discrimination between NL1- and NL2-mediated nuclear localization of the glucocorticoid receptor (1999) Mol. Cell. Biol, 19, pp. 1025-1037
  • Silverstein, A.M., Galigniana, M.D., Kanelakis, K.C., Radanyi, C., Renoir, J.M., Pratt, W.B., Different regions of the immunophilin FKBP52 determine its association with the glucocorticoid receptor, hsp90, and cytoplasmic dynein (1999) J. Biol. Chem, 274, pp. 36980-36986
  • Stewart, M., Molecular mechanism of the nuclear protein import cycle (2007) Nat. Rev. Mol. Cell Biol, 8, pp. 195-208
  • Tanaka, M., Nishi, M., Morimoto, M., Sugimoto, T., Kawata, M., Yellow fluorescent protein-tagged and cyan fluorescent protein-tagged imaging analysis of glucocorticoid receptor and importins in single living cells (2003) Endocrinology, 144, pp. 4070-4079
  • Terry, L.J., Shows, E.B., Wente, S.R., Crossing the nuclear envelope: Hierarchical regulation of nucleocytoplasmic transport (2007) Science, 318, pp. 1412-1416
  • Vicent, G.P., Pecci, A., Ghini, A., Piwien-Pilipuk, G., Galigniana, M.D., Differences in nuclear retention characteristics of agonist-activated glucocorticoid receptor may determine specific responses (2002) Exp. Cell Res, 276, pp. 142-154
  • Witchel, S.F., DeFranco, D.B., Mechanisms of disease: Regulation of glucocorticoid and receptor levels-impact on the metabolic syndrome (2006) Nat. Clin. Pract. Endocrinol. Metab, 2, pp. 621-631
  • Wochnik, G.M., Ruegg, J., Abel, G.A., Schmidt, U., Holsboer, F., Rein, T., FK506-binding proteins 51 and 52 differentially regulate dynein interaction and nuclear translocation of the glucocorticoid receptor in mammalian cells (2005) J. Biol. Chem, 280, pp. 4609-4616


---------- APA ----------
Echeverría, P.C., Mazaira, G., Erlejman, A., Gomez-Sanchez, C., Pilipuk, G.P. & Galigniana, M.D. (2009) . Nuclear import of the glucocorticoid receptor-hsp90 complex through the nuclear pore complex is mediated by its interaction with Nup62 and importin β. Molecular and Cellular Biology, 29(17), 4788-4797.
---------- CHICAGO ----------
Echeverría, P.C., Mazaira, G., Erlejman, A., Gomez-Sanchez, C., Pilipuk, G.P., Galigniana, M.D. "Nuclear import of the glucocorticoid receptor-hsp90 complex through the nuclear pore complex is mediated by its interaction with Nup62 and importin β" . Molecular and Cellular Biology 29, no. 17 (2009) : 4788-4797.
---------- MLA ----------
Echeverría, P.C., Mazaira, G., Erlejman, A., Gomez-Sanchez, C., Pilipuk, G.P., Galigniana, M.D. "Nuclear import of the glucocorticoid receptor-hsp90 complex through the nuclear pore complex is mediated by its interaction with Nup62 and importin β" . Molecular and Cellular Biology, vol. 29, no. 17, 2009, pp. 4788-4797.
---------- VANCOUVER ----------
Echeverría, P.C., Mazaira, G., Erlejman, A., Gomez-Sanchez, C., Pilipuk, G.P., Galigniana, M.D. Nuclear import of the glucocorticoid receptor-hsp90 complex through the nuclear pore complex is mediated by its interaction with Nup62 and importin β. Mol. Cell. Biol. 2009;29(17):4788-4797.