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Abstract:

Rapid, ligand-dependent movement of glucocorticoid receptors (GR) from cytoplasm to the nucleus is hsp90-dependent, and much of the movement system has been defined. GR-hsp90 heterocomplexes isolated from cells contain one of several hsp90-binding immunophilins that link the complex to cytoplasmic dynein, a molecular motor that processes along microtubular tracks to the nucleus. The immunophilins link to dynein indirectly via the dynamitin component of the dynein-associated dynactin complex (Galigniana, M. D., Harrell, J. M., O'Hagen, H. M., Ljungman, M., and Pratt, W. B. (2004) J. Biol. Chem. 279, 22483-22489). Although it is known that rapid, hsp90-dependent GR movement requires intact microtubules, it has not been shown that the movement is dynein-dependent. Here, we show that overexpression of dynamitin, which blocks movement by dissociating the dynein motor from its cargo, inhibits ligand-dependent movement of the GR to the nucleus. We show that native GR·hsp90·immnunophilin complexes contain dynamitin as well as dynein and that GR heterocomplexes isolated from cytosol containing paclitaxel and GTP to stabilize microtubules also contain tubulin. The complete movement system, including the dynein motor complex and tubulin, can be assembled under cell-free conditions by incubating GR immune pellets with paclitaxel/GTP-stabilized cytosol prepared from GR - L cells. This is the first evidence that the movement of a steroid receptor is dynein-dependent, and it is the first isolation of a steroid receptor bound to the entire system that determines its retrograde movement.

Registro:

Documento: Artículo
Título:Evidence for glucocorticoid receptor transport on microtubules by dynein
Autor:Harrell, J.M.; Murphy, P.J.M.; Morishima, Y.; Chen, H.; Mansfield, J.F.; Galigniana, M.D.; Pratt, W.B.
Filiación:Department of Pharmacology, Univ. of Michigan Medical School, Ann Arbor, MI 48109, United States
Univ. of Michigan Sch. of Dentistry, Ann Arbor, MI 48109, United States
University of Michigan, N. Camp. Electron Microbeam Anal. L., Univ. of Michigan Coll. of Eng., Ann Arbor, MI 48109, United States
Departmento de Quimica Biologica, Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires C1428EGA, Argentina
Dept. of Pharmacology, Univ. of Michigan Medical School, 1301 MSRB III, Ann Arbor, MI 48109-0632, United States
Palabras clave:Binding energy; Biochemistry; Immunology; Cytoplasm; Glucocorticoid receptors (GR); Immunophilins; Steroids; Cells; cell protein; dynactin; dynamitin; dynein adenosine triphosphatase; glucocorticoid receptor; guanosine triphosphate; heat shock protein 90; immunophilin; molecular motor; paclitaxel; steroid receptor; tubulin; unclassified drug; animal cell; article; cell nucleus; controlled study; cytoplasm; dissociation; gene overexpression; microtubule; mouse; nonhuman; priority journal; protein assembly; protein transport; Animals; Antibodies, Monoclonal; Binding Sites; Biological Transport; Cell Line; Cell Nucleus; Cytoplasm; Dynein ATPase; Fluorescent Antibody Technique; Gene Expression; Guanosine Triphosphate; HSP90 Heat-Shock Proteins; Immunophilins; Immunosorbent Techniques; Kinetics; Mice; Microscopy, Atomic Force; Microtubule-Associated Proteins; Microtubules; Models, Molecular; NIH 3T3 Cells; Paclitaxel; Rabbits; Rats; Receptors, Glucocorticoid; Transfection; Tubulin; Animalia
Año:2004
Volumen:279
Número:52
Página de inicio:54647
Página de fin:54654
DOI: http://dx.doi.org/10.1074/jbc.M406863200
Título revista:Journal of Biological Chemistry
Título revista abreviado:J. Biol. Chem.
ISSN:00219258
CODEN:JBCHA
CAS:guanosine triphosphate, 86-01-1; paclitaxel, 33069-62-4; Antibodies, Monoclonal; dynactin, 144198-36-7; Dynein ATPase, EC 3.6.4.2; Guanosine Triphosphate, 86-01-1; HSP90 Heat-Shock Proteins; Immunophilins, EC 5.2.1.8; Microtubule-Associated Proteins; Paclitaxel, 33069-62-4; Receptors, Glucocorticoid; Tubulin
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00219258_v279_n52_p54647_Harrell.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v279_n52_p54647_Harrell

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Citas:

---------- APA ----------
Harrell, J.M., Murphy, P.J.M., Morishima, Y., Chen, H., Mansfield, J.F., Galigniana, M.D. & Pratt, W.B. (2004) . Evidence for glucocorticoid receptor transport on microtubules by dynein. Journal of Biological Chemistry, 279(52), 54647-54654.
http://dx.doi.org/10.1074/jbc.M406863200
---------- CHICAGO ----------
Harrell, J.M., Murphy, P.J.M., Morishima, Y., Chen, H., Mansfield, J.F., Galigniana, M.D., et al. "Evidence for glucocorticoid receptor transport on microtubules by dynein" . Journal of Biological Chemistry 279, no. 52 (2004) : 54647-54654.
http://dx.doi.org/10.1074/jbc.M406863200
---------- MLA ----------
Harrell, J.M., Murphy, P.J.M., Morishima, Y., Chen, H., Mansfield, J.F., Galigniana, M.D., et al. "Evidence for glucocorticoid receptor transport on microtubules by dynein" . Journal of Biological Chemistry, vol. 279, no. 52, 2004, pp. 54647-54654.
http://dx.doi.org/10.1074/jbc.M406863200
---------- VANCOUVER ----------
Harrell, J.M., Murphy, P.J.M., Morishima, Y., Chen, H., Mansfield, J.F., Galigniana, M.D., et al. Evidence for glucocorticoid receptor transport on microtubules by dynein. J. Biol. Chem. 2004;279(52):54647-54654.
http://dx.doi.org/10.1074/jbc.M406863200