Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1987 Apr;7(4):1508–1517. doi: 10.1128/mcb.7.4.1508

Glucocorticoid-regulated compartmentalization of cell surface-associated glycoproteins in rat hepatoma cells: evidence for an independent response that requires receptor function and de novo RNA synthesis.

O K Haffar, A K Vallerga, S A Marenda, H J Witchel, G L Firestone
PMCID: PMC365239  PMID: 3037324

Abstract

The role of glucocorticoid hormones in the compartmentalization of cell surface-associated mouse mammary tumor virus (MMTV) glycoproteins was examined in M1.54, a cloned line of MMTV-infected rat hepatoma tissue culture cells. The expression of cellular [2-3H]mannose-labeled and cell surface 125I-labeled MMTV glycoproteins was examined throughout a time course of exposure to dexamethasone, a synthetic glucocorticoid. Posttranslational localization of cell surface MMTV glycoproteins required 6 h of exposure to hormone and occurred approximately 4 h after their initial production in an intracellular fraction. This regulated localization to the cell surface correlated with glucocorticoid receptor occupancy and was inhibited by exposure to RU 38486, a powerful antagonist of glucocorticoid-mediated responses. Cell surface immunoprecipitation demonstrated that actinomycin D, an inhibitor of de novo RNA synthesis, prevented regulated expression of cell surface viral glycoproteins, suggesting that newly synthesized cellular components mediate this process. The localization of cell surface MMTV glycoproteins appeared normal in a transcriptional variant (CR1) that produces basal levels of MMTV RNA and glycoprotein precursors in the presence of dexamethasone. Thus, regulated compartmentalization of viral glycoproteins is not an obligate consequence of a critical precursor concentration. Taken together, our results suggest that posttranslational trafficking of cell surface-destined MMTV glycoproteins resulted from an independent glucocorticoid hormone response that required receptor function and de novo RNA synthesis.

Full text

PDF
1508

Images in this article

1511Image
on p.1511
1512Image
on p.1512
1513Image
on p.1513
1513Image
on p.1513
1514Image
on p.1514
1514Image
on p.1514
1515Image
on p.1515

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bambara R., Wu R. DNA sequence analysis. Terminal sequences of bacteriophage phi80. J Biol Chem. 1975 Jun 25;250(12):4607–4618. [PubMed] [Google Scholar]
  2. Baumann H., Firestone G. L., Burgess T. L., Gross K. W., Yamamoto K. R., Held W. A. Dexamethasone regulation of alpha 1-acid glycoprotein and other acute phase reactants in rat liver and hepatoma cells. J Biol Chem. 1983 Jan 10;258(1):563–570. [PubMed] [Google Scholar]
  3. Baxter J. D., Forsham P. H. Tissue effects of glucocorticoids. Am J Med. 1972 Nov;53(5):573–589. doi: 10.1016/0002-9343(72)90154-4. [DOI] [PubMed] [Google Scholar]
  4. Blobel G. Intracellular protein topogenesis. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1496–1500. doi: 10.1073/pnas.77.3.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Capasso J. M., Abeijon C., Hirschberg C. B. Phosphoproteins and protein kinases of the Golgi apparatus membrane. J Biol Chem. 1985 Nov 25;260(27):14879–14884. [PubMed] [Google Scholar]
  6. Cardiff R. D., Puentes M. J., Young L. J., Smith G. H., Teramoto Y. A., Altrock B. W., Pratt T. S. Serological and biochemical characterization of the mouse mammary tumor virus with localization of p10. Virology. 1978 Mar;85(1):157–167. doi: 10.1016/0042-6822(78)90420-8. [DOI] [PubMed] [Google Scholar]
  7. Chandler V. L., Maler B. A., Yamamoto K. R. DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo. Cell. 1983 Jun;33(2):489–499. doi: 10.1016/0092-8674(83)90430-0. [DOI] [PubMed] [Google Scholar]
  8. Cone R. E., Marchalonis J. J. Surface proteins of thymus-derived lymphocytes and bone-marrow-derived lymphocytes. Selective isolation of immunoglobulin and the theta-antigen by non-ionic detergents. Biochem J. 1974 Jun;140(3):345–354. doi: 10.1042/bj1400345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deutscher S. L., Hirschberg C. B. Mechanism of galactosylation in the Golgi apparatus. A Chinese hamster ovary cell mutant deficient in translocation of UDP-galactose across Golgi vesicle membranes. J Biol Chem. 1986 Jan 5;261(1):96–100. [PubMed] [Google Scholar]
  10. Dickson C., Atterwill M. Structure and processing of the mouse mammary tumor virus glycoprotein precursor pr73env. J Virol. 1980 Aug;35(2):349–361. doi: 10.1128/jvi.35.2.349-361.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dunphy W. G., Rothman J. E. Compartmental organization of the Golgi stack. Cell. 1985 Aug;42(1):13–21. doi: 10.1016/s0092-8674(85)80097-0. [DOI] [PubMed] [Google Scholar]
  12. Firestone G. L., John N. J., Yamamoto K. R. Glucocorticoid-regulated glycoprotein maturation in wild-type and mutant rat cell lines. J Cell Biol. 1986 Dec;103(6 Pt 1):2323–2331. doi: 10.1083/jcb.103.6.2323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Firestone G. L., Payvar F., Yamamoto K. R. Glucocorticoid regulation of protein processing and compartmentalization. Nature. 1982 Nov 18;300(5889):221–225. doi: 10.1038/300221a0. [DOI] [PubMed] [Google Scholar]
  14. Firestone G. L. The role of protein glycosylation in the compartmentalization and processing of mouse mammary tumor virus glycoproteins in mouse mammary tumor virus-infected rat hepatoma cells. J Biol Chem. 1983 May 25;258(10):6155–6161. [PubMed] [Google Scholar]
  15. Firestone G. L., Yamamoto K. R. Two classes of mutant mammary tumor virus-infected HTC cell with defects in glucocorticoid-regulated gene expression. Mol Cell Biol. 1983 Feb;3(2):149–160. doi: 10.1128/mcb.3.2.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fischer S. G. Peptide mapping in gels. Methods Enzymol. 1983;100:424–430. doi: 10.1016/0076-6879(83)00071-3. [DOI] [PubMed] [Google Scholar]
  17. Haffar O. K., Edwards C. P., Firestone G. L. Regulation of alpha 1-acid glycoprotein externalization and intracellular accumulation in glucocorticoid-induced rat hepatoma cells. Arch Biochem Biophys. 1986 Apr;246(1):449–459. doi: 10.1016/0003-9861(86)90491-1. [DOI] [PubMed] [Google Scholar]
  18. Holowka D., Hartmann H., Kanellopoulos J., Metzger H. Association of the receptor for immunoglobulin E with an endogenous polypeptide on rat basophilic leukemia cells. J Recept Res. 1980;1(1):41–68. doi: 10.3109/10799898009039254. [DOI] [PubMed] [Google Scholar]
  19. Ivarie R. D., O'Farrell P. H. The glucocorticoid domain: steroid-mediated changes in the rate of synthesis of rat hepatoma proteins. Cell. 1978 Jan;13(1):41–55. doi: 10.1016/0092-8674(78)90136-8. [DOI] [PubMed] [Google Scholar]
  20. Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
  21. Karlsen K., Vallerga A. K., Hone J., Firestone G. L. A distinct glucocorticoid hormone response regulates phosphoprotein maturation in rat hepatoma cells. Mol Cell Biol. 1986 Feb;6(2):574–585. doi: 10.1128/mcb.6.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kelly R. B. Pathways of protein secretion in eukaryotes. Science. 1985 Oct 4;230(4721):25–32. doi: 10.1126/science.2994224. [DOI] [PubMed] [Google Scholar]
  23. Krangel M. S., Orr H. T., Strominger J. L. Assembly and maturation of HLA-A and HLA-B antigens in vivo. Cell. 1979 Dec;18(4):979–991. doi: 10.1016/0092-8674(79)90210-1. [DOI] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Liu M. C., Yu S., Sy J., Redman C. M., Lipmann F. Tyrosine sulfation of proteins from the human hepatoma cell line HepG2. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7160–7164. doi: 10.1073/pnas.82.21.7160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Logsdon C. D., Moessner J., Williams J. A., Goldfine I. D. Glucocorticoids increase amylase mRNA levels, secretory organelles, and secretion in pancreatic acinar AR42J cells. J Cell Biol. 1985 Apr;100(4):1200–1208. doi: 10.1083/jcb.100.4.1200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moguilewsky M., Philibert D. RU 38486: potent antiglucocorticoid activity correlated with strong binding to the cytosolic glucocorticoid receptor followed by an impaired activation. J Steroid Biochem. 1984 Jan;20(1):271–276. doi: 10.1016/0022-4731(84)90216-4. [DOI] [PubMed] [Google Scholar]
  28. Nelson J. D., Jato-Rodriguez J. J., Labrie F., Mookerjea S. Glycosyltransferase and UDP-galactose pyrophosphatase activities in the endometrium during oestrous cycle of the rat. J Endocrinol. 1977 Apr;73(1):53–58. doi: 10.1677/joe.0.0730053. [DOI] [PubMed] [Google Scholar]
  29. Payvar F., DeFranco D., Firestone G. L., Edgar B., Wrange O., Okret S., Gustafsson J. A., Yamamoto K. R. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell. 1983 Dec;35(2 Pt 1):381–392. doi: 10.1016/0092-8674(83)90171-x. [DOI] [PubMed] [Google Scholar]
  30. Payvar F., Firestone G. L., Ross S. R., Chandler V. L., Wrange O., Carlstedt-Duke J., Gustafsson J. A., Yamamoto K. R. Multiple specific binding sites for purified glucocorticoid receptors on mammary tumor virus DNA. J Cell Biochem. 1982;19(3):241–247. doi: 10.1002/jcb.240190305. [DOI] [PubMed] [Google Scholar]
  31. Payvar F., Wrange O., Carlstedt-Duke J., Okret S., Gustafsson J. A., Yamamoto K. R. Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6628–6632. doi: 10.1073/pnas.78.11.6628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Platt E. J., Karlsen K., Lopez-Valdivieso A., Cook P. W., Firestone G. L. Highly sensitive immunoadsorption procedure for detection of low-abundance proteins. Anal Biochem. 1986 Jul;156(1):126–135. doi: 10.1016/0003-2697(86)90163-6. [DOI] [PubMed] [Google Scholar]
  33. Ponta H., Kennedy N., Skroch P., Hynes N. E., Groner B. Hormonal response region in the mouse mammary tumor virus long terminal repeat can be dissociated from the proviral promoter and has enhancer properties. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1020–1024. doi: 10.1073/pnas.82.4.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Redmond S. M., Dickson C. Sequence and expression of the mouse mammary tumour virus env gene. EMBO J. 1983;2(1):125–131. doi: 10.1002/j.1460-2075.1983.tb01393.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ringold G. M., Shank P. R., Varmus H. E., Ring J., Yamamoto K. R. Integration and transcription of mouse mammary tumor virus DNA in rat hepatoma cells. Proc Natl Acad Sci U S A. 1979 Feb;76(2):665–669. doi: 10.1073/pnas.76.2.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sarkar N. H., Racevskis J. Expression and disposition of the murine mammary tumor virus (MuMTV) envelope gene products by murine mammary tumor cells. Virology. 1983 Apr 15;126(1):279–300. doi: 10.1016/0042-6822(83)90479-8. [DOI] [PubMed] [Google Scholar]
  37. Shepherd V. L., Konish M. G., Stahl P. Dexamethasone increases expression of mannose receptors and decreases extracellular lysosomal enzyme accumulation in macrophages. J Biol Chem. 1985 Jan 10;260(1):160–164. [PubMed] [Google Scholar]
  38. Starr C. M., Lucas J. J. Regulation of dolichyl phosphate-mediated protein glycosylation: estrogen effects on glucosyl transfers in oviduct membranes. Arch Biochem Biophys. 1985 Feb 15;237(1):261–270. doi: 10.1016/0003-9861(85)90277-2. [DOI] [PubMed] [Google Scholar]
  39. Ucker D. S., Firestone G. L., Yamamoto K. R. Glucocorticoids and chromosomal position modulate murine mammary tumor virus transcription by affecting efficiency of promoter utilization. Mol Cell Biol. 1983 Apr;3(4):551–561. doi: 10.1128/mcb.3.4.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ucker D. S., Ross S. R., Yamamoto K. R. Mammary tumor virus DNA contains sequences required for its hormone-regulated transcription. Cell. 1981 Dec;27(2 Pt 1):257–266. doi: 10.1016/0092-8674(81)90409-8. [DOI] [PubMed] [Google Scholar]
  41. Wold F. In vivo chemical modification of proteins (post-translational modification). Annu Rev Biochem. 1981;50:783–814. doi: 10.1146/annurev.bi.50.070181.004031. [DOI] [PubMed] [Google Scholar]
  42. Yamamoto K. R. Steroid receptor regulated transcription of specific genes and gene networks. Annu Rev Genet. 1985;19:209–252. doi: 10.1146/annurev.ge.19.120185.001233. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES