Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1983 Feb;3(2):149–160. doi: 10.1128/mcb.3.2.149

Two classes of mutant mammary tumor virus-infected HTC cell with defects in glucocorticoid-regulated gene expression.

G L Firestone, K R Yamamoto
PMCID: PMC368517  PMID: 6300655

Abstract

We have isolated mutant derivatives of M1.54 (a mammary tumor virus [MTV]-infected rat hepatoma [HTC] cell line containing multiple integrated proviruses) that fail to express hormone-inducible cell surface viral glycoproteins. In wild-type M1.54, the synthetic glucocorticoid dexamethasone selectively stimulates the rate of synthesis of MTV RNA. In addition, dexamethasone is essential for posttranslational maturation of three of the four cell surface viral glycoproteins processed from the MTV glycosylated precursor polyprotein; the fourth mature species is produced constitutively. Two mutant phenotypes are described; each contains glucocorticoid receptors that are indistinguishable from the wild-type receptor with respect to hormone affinity, intracellular concentration, nuclear translocation efficiency, DNA-cellulose chromatography, and sedimentation rate. In one class, represented by the mutant line CR1, dexamethasone fails to stimulate the low basal rate of MTV gene transcription; surprisingly, hormonal regulation of tyrosine aminotransferase activity is also defective in CR1, whereas several other cellular responses to dexamethasone are normal. In the second class of mutants, represented by CR4, dexamethasone stimulates synthesis of MTV transcripts indistinguishable from those produced in M1.54, but only the constitutive cell surface viral glycoprotein is expressed. Thus, these mutants define two distinct and novel aspects of glucocorticoid regulated gene expression in HTC cells: CR4 contains a defect in a hormone inducible protein maturation pathway that acts on specific viral (and presumably cellular) precursor polypeptides, whereas the lesion in CR1 appears to affect the expression of a subset of the gene products normally under glucocorticoid control in M1.54.

Full text

PDF
149

Images in this article

153Image
on p.153
154Image
on p.154
156Image
on p.156
157Image
on p.157
158Image
on p.158

Selected References

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

  1. Baumann H., Held W. A. Biosynthesis and hormone-regulated expression of secretory glycoproteins in rat liver and hepatoma cells. Effect of glucocorticoids and inflammation. J Biol Chem. 1981 Oct 10;256(19):10145–10155. [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Carlstedt-Duke J., Okret S., Wrange O., Gustafsson J. A. Immunochemical analysis of the glucocorticoid receptor: identification of a third domain separate from the steroid-binding and DNA-binding domains. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4260–4264. doi: 10.1073/pnas.79.14.4260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Feinstein S. C., Ross S. R., Yamamoto K. R. Chromosomal position effects determine transcriptional potential of integrated mammary tumor virus DNA. J Mol Biol. 1982 Apr 15;156(3):549–565. doi: 10.1016/0022-2836(82)90266-2. [DOI] [PubMed] [Google Scholar]
  8. Firestone G. L., Heath E. C. The cyclic AMP-mediated induction of alkaline phosphatase in mouse L-cells. J Biol Chem. 1981 Feb 10;256(3):1396–1403. [PubMed] [Google Scholar]
  9. 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]
  10. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  11. Grove J. R., Dieckmann B. S., Schroer T. A., Ringold G. M. Isolation of glucocorticoid-unresponsive rat hepatoma cells by fluorescence-activated cell sorting. Cell. 1980 Aug;21(1):47–56. doi: 10.1016/0092-8674(80)90113-0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Hunt T., Jackson R. J. The rabbit reticulocyte lysate as a system for studying mRNA. Hamatol Bluttransfus. 1974;14:300–307. [PubMed] [Google Scholar]
  14. 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]
  15. Jackson V., Chalkley R. The binding of estradiol-17 beta to the bovine endometrial nuclear membrane. J Biol Chem. 1974 Mar 10;249(5):1615–1626. [PubMed] [Google Scholar]
  16. Kabat D., Ruta M., Murray M. J., Polonoff E. Immunoselection of mutants deficient in cell surface glycoproteins encoded by murine erythroleukemia viruses. Proc Natl Acad Sci U S A. 1980 Jan;77(1):57–61. doi: 10.1073/pnas.77.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaslow H. R., Cox D., Groppi V. E., Bourne H. R. An Mr = 52,000 peptide can mediate effects on cholera toxin on adenylate cyclase in intact cells. Mol Pharmacol. 1981 May;19(3):406–410. [PubMed] [Google Scholar]
  18. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. 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]
  21. Liao S., Liang T., Tymoczko J. L. Ribonucleoprotein binding of steroid-"receptor" complexes. Nat New Biol. 1973 Feb 14;241(111):211–213. doi: 10.1038/newbio241211a0. [DOI] [PubMed] [Google Scholar]
  22. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  25. Pfahl M., Kelleher R. J., Jr, Bourgeois S. General features of steroid resistance on lymphoid cell lines. Mol Cell Endocrinol. 1978 Apr;10(2):193–207. doi: 10.1016/0303-7207(78)90125-9. [DOI] [PubMed] [Google Scholar]
  26. Polsky F., Edgell M. H., Seidman J. G., Leder P. High capacity gel preparative electrophoresis for purification of fragments of genomic DNA. Anal Biochem. 1978 Jul 1;87(2):397–410. doi: 10.1016/0003-2697(78)90689-9. [DOI] [PubMed] [Google Scholar]
  27. Ringold G. M. Glucocorticoid regulation of mouse mammary tumor virus gene expression. Biochim Biophys Acta. 1979 Dec 19;560(4):487–508. doi: 10.1016/0304-419x(79)90014-3. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Ringold G. M., Yamamoto K. R., Bishop J. M., Varmus H. E. Glucocorticoid-stimulated accumulation of mouse mammary tumor virus RNA: increased rate of synthesis of viral RNA. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2879–2883. doi: 10.1073/pnas.74.7.2879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ringold G. M., Yamamoto K. R., Tomkins G. M., Bishop M., Varmus H. E. Dexamethasone-mediated induction of mouse mammary tumor virus RNA: a system for studying glucocorticoid action. Cell. 1975 Nov;6(3):299–305. doi: 10.1016/0092-8674(75)90181-6. [DOI] [PubMed] [Google Scholar]
  31. Rousseau G. G., Baxter J. D., Higgins S. J., Tomkins G. M. Steroid-induced nuclear binding of glucocorticoid receptors in intact hepatoma cells. J Mol Biol. 1973 Sep 25;79(3):539–554. doi: 10.1016/0022-2836(73)90405-1. [DOI] [PubMed] [Google Scholar]
  32. Samuels H. H., Tomkins G. M. Relation of steroid structure to enzyme induction in hepatoma tissue culture cells. J Mol Biol. 1970 Aug 28;52(1):57–74. doi: 10.1016/0022-2836(70)90177-4. [DOI] [PubMed] [Google Scholar]
  33. Scherer G., Schmid W., Strange C. M., Röwekamp W., Schütz G. Isolation of cDNA clones coding for rat tyrosine aminotransferase. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7205–7208. doi: 10.1073/pnas.79.23.7205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shank P. R., Cohen J. C., Varmus H. E., Yamamoto K. R., Ringold G. M. Mapping of linear and circular forms of mouse mammary tumor virus DNA with restriction endonucleases: evidence for a large specific deletion occurring at high frequency during circularization. Proc Natl Acad Sci U S A. 1978 May;75(5):2112–2116. doi: 10.1073/pnas.75.5.2112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Szego C. M. The lysosome as a mediator of hormone action. Recent Prog Horm Res. 1974;30(0):171–233. doi: 10.1016/b978-0-12-571130-2.50009-2. [DOI] [PubMed] [Google Scholar]
  37. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Thompson E. B., Granner D. K., Gelehrter T., Erickson J., Hager G. L. Unlinked control of multiple glucocorticoid-induced processes in HTC cells. Mol Cell Endocrinol. 1979 Sep;15(3):135–150. doi: 10.1016/0303-7207(79)90034-0. [DOI] [PubMed] [Google Scholar]
  39. Thompson E. B., Tomkins G. M., Curran J. F. Induction of tyrosine alpha-ketoglutarate transaminase by steroid hormones in a newly established tissue culture cell line. Proc Natl Acad Sci U S A. 1966 Jul;56(1):296–303. doi: 10.1073/pnas.56.1.296. [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. Varmus H. E., Quintrell N., Medeiros E., Bishop J. M., Nowinski R. C., Sarkar N. H. Transcription of mouse mammary tumor virus genes in tissues from high and low tumor incidence mouse strains. J Mol Biol. 1973 Oct 5;79(4):663–679. doi: 10.1016/0022-2836(73)90070-3. [DOI] [PubMed] [Google Scholar]
  42. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  43. Yamamoto K. R., Alberts B. M. Steroid receptors: elements for modulation of eukaryotic transcription. Annu Rev Biochem. 1976;45:721–746. doi: 10.1146/annurev.bi.45.070176.003445. [DOI] [PubMed] [Google Scholar]
  44. Yamamoto K. R., Chandler V. L., Ross S. R., Ucker D. S., Ring J. C., Feinstein S. C. Integration and activity of mammary tumor virus genes: regulation by hormone receptors and chromosomal position. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):687–697. doi: 10.1101/sqb.1981.045.01.086. [DOI] [PubMed] [Google Scholar]
  45. Yamamoto K. R., Gehring U., Stampfer M. R., Sibley C. H. Genetic approaches to steroid hormone action. Recent Prog Horm Res. 1976;32:3–32. doi: 10.1016/b978-0-12-571132-6.50008-7. [DOI] [PubMed] [Google Scholar]
  46. Yamamoto K. R., Stallcup M. R., Ring J., Ringold G. M. Mammary tumor virus DNA: a glucocorticoid-responsive transposable element. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):625–638. doi: 10.1101/sqb.1978.042.01.065. [DOI] [PubMed] [Google Scholar]
  47. Yamamoto K. R., Stampfer M. R., Tomkins G. M. Receptors from glucocorticoid-sensitive lymphoma cells and two clases of insensitive clones: physical and DNA-binding properties. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3901–3905. doi: 10.1073/pnas.71.10.3901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Young H. A., Shih T. Y., Scolnick E. M., Parks W. P. Steroid induction of mouse mammary tumor virus: effect upon synthesis and degradation of viral RNA. J Virol. 1977 Jan;21(1):139–146. doi: 10.1128/jvi.21.1.139-146.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES