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. 1992 Oct 2;119(2):337–348. doi: 10.1083/jcb.119.2.337

The cytoskeleton and the cellular traffic of the progesterone receptor

PMCID: PMC2289648  PMID: 1400578

Abstract

Previous studies on glucocorticoid receptors have suggested the existence of interactions between the receptor and microtubule or actin networks. It was hypothesized that such interactions may contribute to the guidance of steroid hormone receptors towards the nucleus. We used a permanent L cell line expressing the delta 638-642 progesterone receptor. This mutant has all the characteristics of the wild type receptor except that the deletion of five amino acids inactivates the constitutive karyophilic signal. Consequently, the receptor is cytoplasmic in the absence of hormone but is shifted into the nucleus when administration of hormone activates the second karyophilic signal. Optical microscopy and confocal laser microscopy were used in intact cells or in cells depleted of soluble elements by permeabilization with detergents. By immunofluorescence, the receptor was found to be mainly concentrated in the perinuclear area. A small fraction of progesterone receptor (PR) persisted in this region after Triton X100 treatment. These observations suggested that the receptor could interact with some insoluble constituent(s) of the cytoplasm. However, careful colocalization studies showed that this heterogenous distribution was not due to interactions with microtubule, microfilament, or intermediate filament networks. Functional involvement of these networks in the translocation of the receptor into the nucleus was studied after cell treatment with cytoskeletal drugs such as nocodazole, demecolcine and cytochalasin. None of these compounds prevented or even delayed the hormone-dependent transfer of delta 638- 642 PR into the nucleus. Similar conclusions were reached with the wild type receptor expressed by transfection in Cos-7 cells. PR was shifted from the nucleus into the cytoplasm by administration of energy- depleting drugs. After disruption of the various cytoskeletal networks normal nuclear reaccumulation of the receptor was observed when these drugs were removed. The results thus suggest that the progesterone receptor is not colocalized with the main cytoskeletal components. Disruption of the cytoskeletal networks does not prevent its nuclear translocation. Thus, karyophilic signals and interactions with the nuclear pore seem to be the primary determinants of the cellular traffic of the progesterone receptor.

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Selected References

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  1. Adam S. A., Gerace L. Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import. Cell. 1991 Sep 6;66(5):837–847. doi: 10.1016/0092-8674(91)90431-w. [DOI] [PubMed] [Google Scholar]
  2. Akner G., Sundqvist K. G., Denis M., Wikström A. C., Gustafsson J. A. Immunocytochemical localization of glucocorticoid receptor in human gingival fibroblasts and evidence for a colocalization of glucocorticoid receptor with cytoplasmic microtubules. Eur J Cell Biol. 1990 Dec;53(2):390–401. [PubMed] [Google Scholar]
  3. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  4. Breeuwer M., Goldfarb D. S. Facilitated nuclear transport of histone H1 and other small nucleophilic proteins. Cell. 1990 Mar 23;60(6):999–1008. doi: 10.1016/0092-8674(90)90348-i. [DOI] [PubMed] [Google Scholar]
  5. Collier N. C., Schlesinger M. J. The dynamic state of heat shock proteins in chicken embryo fibroblasts. J Cell Biol. 1986 Oct;103(4):1495–1507. doi: 10.1083/jcb.103.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Brabander M. J., Van de Veire R. M., Aerts F. E., Borgers M., Janssen P. A. The effects of methyl (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl) carbamate, (R 17934; NSC 238159), a new synthetic antitumoral drug interfering with microtubules, on mammalian cells cultured in vitro. Cancer Res. 1976 Mar;36(3):905–916. [PubMed] [Google Scholar]
  8. Eckert B. S. Alteration of intermediate filament distribution in PtK1 cells by acrylamide. Eur J Cell Biol. 1985 May;37:169–174. [PubMed] [Google Scholar]
  9. Farman N., Oblin M. E., Lombes M., Delahaye F., Westphal H. M., Bonvalet J. P., Gasc J. M. Immunolocalization of gluco- and mineralocorticoid receptors in rabbit kidney. Am J Physiol. 1991 Feb;260(2 Pt 1):C226–C233. doi: 10.1152/ajpcell.1991.260.2.C226. [DOI] [PubMed] [Google Scholar]
  10. Fulton A. B., Wan K. M., Penman S. The spatial distribution of polyribosomes in 3T3 cells and the associated assembly of proteins into the skeletal framework. Cell. 1980 Jul;20(3):849–857. doi: 10.1016/0092-8674(80)90331-1. [DOI] [PubMed] [Google Scholar]
  11. Fuxe K., Wikström A. C., Okret S., Agnati L. F., Härfstrand A., Yu Z. Y., Granholm L., Zoli M., Vale W., Gustafsson J. A. Mapping of glucocorticoid receptor immunoreactive neurons in the rat tel- and diencephalon using a monoclonal antibody against rat liver glucocorticoid receptor. Endocrinology. 1985 Nov;117(5):1803–1812. doi: 10.1210/endo-117-5-1803. [DOI] [PubMed] [Google Scholar]
  12. Gorski J., Toft D., Shyamala G., Smith D., Notides A. Hormone receptors: studies on the interaction of estrogen with the uterus. Recent Prog Horm Res. 1968;24:45–80. doi: 10.1016/b978-1-4831-9827-9.50008-3. [DOI] [PubMed] [Google Scholar]
  13. Guiochon-Mantel A., Lescop P., Christin-Maitre S., Loosfelt H., Perrot-Applanat M., Milgrom E. Nucleocytoplasmic shuttling of the progesterone receptor. EMBO J. 1991 Dec;10(12):3851–3859. doi: 10.1002/j.1460-2075.1991.tb04954.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Guiochon-Mantel A., Loosfelt H., Lescop P., Sar S., Atger M., Perrot-Applanat M., Milgrom E. Mechanisms of nuclear localization of the progesterone receptor: evidence for interaction between monomers. Cell. 1989 Jun 30;57(7):1147–1154. doi: 10.1016/0092-8674(89)90052-4. [DOI] [PubMed] [Google Scholar]
  15. Husmann D. A., Wilson C. M., McPhaul M. J., Tilley W. D., Wilson J. D. Antipeptide antibodies to two distinct regions of the androgen receptor localize the receptor protein to the nuclei of target cells in the rat and human prostate. Endocrinology. 1990 May;126(5):2359–2368. doi: 10.1210/endo-126-5-2359. [DOI] [PubMed] [Google Scholar]
  16. Hynes R. O., Destree A. T. 10 nm filaments in normal and transformed cells. Cell. 1978 Jan;13(1):151–163. doi: 10.1016/0092-8674(78)90146-0. [DOI] [PubMed] [Google Scholar]
  17. Jensen E. V., Suzuki T., Kawashima T., Stumpf W. E., Jungblut P. W., DeSombre E. R. A two-step mechanism for the interaction of estradiol with rat uterus. Proc Natl Acad Sci U S A. 1968 Feb;59(2):632–638. doi: 10.1073/pnas.59.2.632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. A short amino acid sequence able to specify nuclear location. Cell. 1984 Dec;39(3 Pt 2):499–509. doi: 10.1016/0092-8674(84)90457-4. [DOI] [PubMed] [Google Scholar]
  19. King W. J., Greene G. L. Monoclonal antibodies localize oestrogen receptor in the nuclei of target cells. Nature. 1984 Feb 23;307(5953):745–747. doi: 10.1038/307745a0. [DOI] [PubMed] [Google Scholar]
  20. Koyasu S., Nishida E., Kadowaki T., Matsuzaki F., Iida K., Harada F., Kasuga M., Sakai H., Yahara I. Two mammalian heat shock proteins, HSP90 and HSP100, are actin-binding proteins. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8054–8058. doi: 10.1073/pnas.83.21.8054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. LaCasse E. C., Lefebvre Y. A. Nuclear and nuclear envelope binding proteins of the glucocorticoid receptor nuclear localization peptide identified by crosslinking. J Steroid Biochem Mol Biol. 1991;40(1-3):279–285. doi: 10.1016/0960-0760(91)90193-9. [DOI] [PubMed] [Google Scholar]
  22. Lanford R. E., Kanda P., Kennedy R. C. Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell. 1986 Aug 15;46(4):575–582. doi: 10.1016/0092-8674(86)90883-4. [DOI] [PubMed] [Google Scholar]
  23. Lehto V. P., Virtanen I., Kurki P. Intermediate filaments anchor the nuclei in nuclear monolayers of cultured human fibroblasts. Nature. 1978 Mar 9;272(5649):175–177. doi: 10.1038/272175a0. [DOI] [PubMed] [Google Scholar]
  24. Logeat F., Vu Hai M. T., Fournier A., Legrain P., Buttin G., Milgrom E. Monoclonal antibodies to rabbit progesterone receptor: crossreaction with other mammalian progesterone receptors. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6456–6459. doi: 10.1073/pnas.80.21.6456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lombès M., Farman N., Oblin M. E., Baulieu E. E., Bonvalet J. P., Erlanger B. F., Gasc J. M. Immunohistochemical localization of renal mineralocorticoid receptor by using an anti-idiotypic antibody that is an internal image of aldosterone. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1086–1088. doi: 10.1073/pnas.87.3.1086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lorenzo F., Jolivet A., Loosfelt H., Thu vu Hai M., Brailly S., Perrot-Applanat M., Milgrom E. A rapid method of epitope mapping. Application to the study of immunogenic domains and to the characterization of various forms of rabbit progesterone receptor. Eur J Biochem. 1988 Sep 1;176(1):53–60. doi: 10.1111/j.1432-1033.1988.tb14250.x. [DOI] [PubMed] [Google Scholar]
  27. Miyata Y., Yahara I. Cytoplasmic 8 S glucocorticoid receptor binds to actin filaments through the 90-kDa heat shock protein moiety. J Biol Chem. 1991 May 15;266(14):8779–8783. [PubMed] [Google Scholar]
  28. Osborn M., Weber K. The display of microtubules in transformed cells. Cell. 1977 Nov;12(3):561–571. doi: 10.1016/0092-8674(77)90257-4. [DOI] [PubMed] [Google Scholar]
  29. Perrot-Applanat M., Groyer-Picard M. T., Lorenzo F., Jolivet A., Vu Hai M. T., Pallud C., Spyratos F., Milgrom E. Immunocytochemical study with monoclonal antibodies to progesterone receptor in human breast tumors. Cancer Res. 1987 May 15;47(10):2652–2661. [PubMed] [Google Scholar]
  30. Perrot-Applanat M., Logeat F., Groyer-Picard M. T., Milgrom E. Immunocytochemical study of mammalian progesterone receptor using monoclonal antibodies. Endocrinology. 1985 Apr;116(4):1473–1484. doi: 10.1210/endo-116-4-1473. [DOI] [PubMed] [Google Scholar]
  31. Picard D., Kumar V., Chambon P., Yamamoto K. R. Signal transduction by steroid hormones: nuclear localization is differentially regulated in estrogen and glucocorticoid receptors. Cell Regul. 1990 Feb;1(3):291–299. doi: 10.1091/mbc.1.3.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Picard D., Yamamoto K. R. Two signals mediate hormone-dependent nuclear localization of the glucocorticoid receptor. EMBO J. 1987 Nov;6(11):3333–3340. doi: 10.1002/j.1460-2075.1987.tb02654.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pratt W. B. Glucocorticoid receptor structure and the initial events in signal transduction. Prog Clin Biol Res. 1990;322:119–132. [PubMed] [Google Scholar]
  34. Renoir J. M., Radanyi C., Faber L. E., Baulieu E. E. The non-DNA-binding heterooligomeric form of mammalian steroid hormone receptors contains a hsp90-bound 59-kilodalton protein. J Biol Chem. 1990 Jun 25;265(18):10740–10745. [PubMed] [Google Scholar]
  35. Richardson W. D., Mills A. D., Dilworth S. M., Laskey R. A., Dingwall C. Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores. Cell. 1988 Mar 11;52(5):655–664. doi: 10.1016/0092-8674(88)90403-5. [DOI] [PubMed] [Google Scholar]
  36. Sanchez E. R., Faber L. E., Henzel W. J., Pratt W. B. The 56-59-kilodalton protein identified in untransformed steroid receptor complexes is a unique protein that exists in cytosol in a complex with both the 70- and 90-kilodalton heat shock proteins. Biochemistry. 1990 May 29;29(21):5145–5152. doi: 10.1021/bi00473a021. [DOI] [PubMed] [Google Scholar]
  37. Sanchez E. R., Redmond T., Scherrer L. C., Bresnick E. H., Welsh M. J., Pratt W. B. Evidence that the 90-kilodalton heat shock protein is associated with tubulin-containing complexes in L cell cytosol and in intact PtK cells. Mol Endocrinol. 1988 Aug;2(8):756–760. doi: 10.1210/mend-2-8-756. [DOI] [PubMed] [Google Scholar]
  38. Scherrer L. C., Dalman F. C., Massa E., Meshinchi S., Pratt W. B. Structural and functional reconstitution of the glucocorticoid receptor-hsp90 complex. J Biol Chem. 1990 Dec 15;265(35):21397–21400. [PubMed] [Google Scholar]
  39. Schliwa M. Action of cytochalasin D on cytoskeletal networks. J Cell Biol. 1982 Jan;92(1):79–91. doi: 10.1083/jcb.92.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Silver P. A. How proteins enter the nucleus. Cell. 1991 Feb 8;64(3):489–497. doi: 10.1016/0092-8674(91)90233-o. [DOI] [PubMed] [Google Scholar]
  41. Tai P. K., Maeda Y., Nakao K., Wakim N. G., Duhring J. L., Faber L. E. A 59-kilodalton protein associated with progestin, estrogen, androgen, and glucocorticoid receptors. Biochemistry. 1986 Sep 9;25(18):5269–5275. doi: 10.1021/bi00366a043. [DOI] [PubMed] [Google Scholar]
  42. Welch W. J., Feramisco J. R. Disruption of the three cytoskeletal networks in mammalian cells does not affect transcription, translation, or protein translocation changes induced by heat shock. Mol Cell Biol. 1985 Jul;5(7):1571–1581. doi: 10.1128/mcb.5.7.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Welshons W. V., Lieberman M. E., Gorski J. Nuclear localization of unoccupied oestrogen receptors. Nature. 1984 Feb 23;307(5953):747–749. doi: 10.1038/307747a0. [DOI] [PubMed] [Google Scholar]
  44. Wikström A. C., Bakke O., Okret S., Brönnegård M., Gustafsson J. A. Intracellular localization of the glucocorticoid receptor: evidence for cytoplasmic and nuclear localization. Endocrinology. 1987 Apr;120(4):1232–1242. doi: 10.1210/endo-120-4-1232. [DOI] [PubMed] [Google Scholar]

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