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. 1971 Nov;125(1):285–295. doi: 10.1042/bj1250285

A reconstituted cell-free system for the specific transfer of steroid–receptor complexes into nuclear chromatin isolated from rat ventral prostate gland

W I P Mainwaring 1, Brenda M Peterken 1
PMCID: PMC1178052  PMID: 5158914

Abstract

1. A system has been developed for the specific transfer of [3H]dihydrotestosterone–receptor complexes into prostatic chromatin in vitro. 2. Under optimum conditions the overall transfer of [3H]dihydrotestosterone into purified chromatin in this reconstituted system is entirely consistent with the results obtained in whole tissue both in vivo and in vitro. 3. The transfer of [3H]dihydrotestosterone into chromatin is tissue-specific and maximal into chromatin isolated from androgen-dependent tissues. 4. The tissue specificity is maintained at two levels: first, in the presence of specific cytoplasmic androgen-receptor proteins; secondly, by the nature and composition of the chromatin itself. 5. Evidence is presented that androgenic steroids in vivo may maintain the tissue-specific nature of chromatin in androgen-dependent tissues by the selective induction of nuclear protein synthesis. 6. The relevance of these findings to the mechanism of action of androgenic steroids is discussed.

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Selected References

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

  1. Alberts B. M., Amodio F. J., Jenkins M., Gutmann E. D., Ferris F. L. Studies with DNA-cellulose chromatography. I. DNA-binding proteins from Escherichia coli. Cold Spring Harb Symp Quant Biol. 1968;33:289–305. doi: 10.1101/sqb.1968.033.01.033. [DOI] [PubMed] [Google Scholar]
  2. Anderson K. M., Liao S. Selective retention of dihydrotestosterone by prostatic nuclei. Nature. 1968 Jul 20;219(5151):277–279. doi: 10.1038/219277a0. [DOI] [PubMed] [Google Scholar]
  3. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bekhor I., Kung G. M., Bonner J. Sequence-specific interaction of DNA and chromosomal protein. J Mol Biol. 1969 Jan;39(2):351–364. doi: 10.1016/0022-2836(69)90322-2. [DOI] [PubMed] [Google Scholar]
  5. Bruchovsky N., Wilson J. D. The conversion of testosterone to 5-alpha-androstan-17-beta-ol-3-one by rat prostate in vivo and in vitro. J Biol Chem. 1968 Apr 25;243(8):2012–2021. [PubMed] [Google Scholar]
  6. Clark R. J., Felsenfeld G. Structure of chromatin. Nat New Biol. 1971 Jan 27;229(4):101–106. doi: 10.1038/newbio229101a0. [DOI] [PubMed] [Google Scholar]
  7. Coffey D. S., Shimazaki J., Williams-Ashman H. G. Polymerization of deoxyribonucleotides in relation to androgen-induced prostatic growth. Arch Biochem Biophys. 1968 Mar 20;124(1):184–198. doi: 10.1016/0003-9861(68)90319-6. [DOI] [PubMed] [Google Scholar]
  8. Edwards L. J., Hnilica L. S. The specificity of histones in nucleated erythrocytes. Experientia. 1968 Mar 15;24(3):228–229. doi: 10.1007/BF02152784. [DOI] [PubMed] [Google Scholar]
  9. FARNSWORTH W. E., BROWN J. R. TESTOSTERONE METABOLISM IN THE PROSTATE. Natl Cancer Inst Monogr. 1963 Oct;12:323–329. [PubMed] [Google Scholar]
  10. Gilbert W., Müller-Hill B. Isolation of the lac repressor. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1891–1898. doi: 10.1073/pnas.56.6.1891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gurdon J. B., Woodland H. R. The cytoplasmic control of nuclear activity in animal development. Biol Rev Camb Philos Soc. 1968 May;43(2):233–267. doi: 10.1111/j.1469-185x.1968.tb00960.x. [DOI] [PubMed] [Google Scholar]
  12. HYMER W. C., KUFF E. L. ISOLATION OF NUCLEI FROM MAMMALIAN TISSUES THROUGH THE USE OF TRITON X-100. J Histochem Cytochem. 1964 May;12:359–363. doi: 10.1177/12.5.359. [DOI] [PubMed] [Google Scholar]
  13. Hardin J. W., O'Brien T. J., Cherry J. H. Stimulation of chromatin-bound RNA polymerase activity by a soluble factor. Biochim Biophys Acta. 1970 Dec 14;224(2):667–670. doi: 10.1016/0005-2787(70)90609-x. [DOI] [PubMed] [Google Scholar]
  14. Itzhaki R. F. Studies on the accessibility of deoxyribonucleic acid in deoxyribonucleoprotein to cationic molecules. Biochem J. 1971 May;122(4):583–592. doi: 10.1042/bj1220583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
  16. KREBS H. A. Body size and tissue respiration. Biochim Biophys Acta. 1950 Jan;4(1-3):249–269. doi: 10.1016/0006-3002(50)90032-1. [DOI] [PubMed] [Google Scholar]
  17. Kleinsmith L. J., Heidema J., Carroll A. Specific binding of rat liver nuclear proteins to DNA. Nature. 1970 Jun 13;226(5250):1025–1026. doi: 10.1038/2261025a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Mainwaring W. I. A soluble androgen receptor in the cytoplasm of rat prostate. J Endocrinol. 1969 Dec;45(4):531–541. doi: 10.1677/joe.0.0450531. [DOI] [PubMed] [Google Scholar]
  20. Mainwaring W. I. Changes in the ribonucleic acid metabolism of aging mouse tissues with particular reference to the prostate gland. Biochem J. 1968 Nov;110(1):79–86. doi: 10.1042/bj1100079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mainwaring W. I., Mangan F. R., Peterken B. M. Studies on the solubilized ribonucleic acid polymerase from rat ventral prostate gland. Biochem J. 1971 Jul;123(4):619–628. doi: 10.1042/bj1230619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mainwaring W. I. The binding of (1,2-3H)testosterone within nuclei of the rat prostate. J Endocrinol. 1969 Jul;44(3):323–333. doi: 10.1677/joe.0.0440323. [DOI] [PubMed] [Google Scholar]
  23. Marushige K., Bonner J. Template properties of liver chromatin. J Mol Biol. 1966 Jan;15(1):160–174. doi: 10.1016/s0022-2836(66)80218-8. [DOI] [PubMed] [Google Scholar]
  24. Matthysse A. G. Organ specificity of hormone-receptor-chromatin interactions. Biochim Biophys Acta. 1970 Feb 18;199(2):519–521. doi: 10.1016/0005-2787(70)90094-8. [DOI] [PubMed] [Google Scholar]
  25. Merriam R. W. Movement of cytoplasmic proteins into nuclei induced to enlarge and initiate DNA or RNA synthesis. J Cell Sci. 1969 Sep;5(2):333–349. doi: 10.1242/jcs.5.2.333. [DOI] [PubMed] [Google Scholar]
  26. Ofner P. Effects and metabolism of hormones in normal and neoplastic prostate tissue. Vitam Horm. 1968;26:237–291. doi: 10.1016/s0083-6729(08)60756-6. [DOI] [PubMed] [Google Scholar]
  27. Panyim S., Chalkley R. High resolution acrylamide gel electrophoresis of histones. Arch Biochem Biophys. 1969 Mar;130(1):337–346. doi: 10.1016/0003-9861(69)90042-3. [DOI] [PubMed] [Google Scholar]
  28. Ptashne M. Specific binding of the lambda phage repressor to lambda DNA. Nature. 1967 Apr 15;214(5085):232–234. doi: 10.1038/214232a0. [DOI] [PubMed] [Google Scholar]
  29. Salas J., Green H. Proteins binding to DNA and their relation to growth in cultured mammalian cells. Nat New Biol. 1971 Feb 10;229(6):165–169. doi: 10.1038/newbio229165a0. [DOI] [PubMed] [Google Scholar]
  30. Shelton K. R., Allfrey V. G. Selective synthesis of a nuclear acidic protein in liver cells stimulated by cortisol. Nature. 1970 Oct 10;228(5267):132–134. doi: 10.1038/228132a0. [DOI] [PubMed] [Google Scholar]
  31. Steggles A. W., Spelsberg T. C., Glasser S. R., O'Malley B. W. Soluble complexes between steroid hormones and target-tissue receptors bind specifically to target-tissue chromatin. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1479–1482. doi: 10.1073/pnas.68.7.1479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Swaneck G. E., Chu L. L., Edelman I. S. Stereospecific binding of aldosterone to renal chromatin. J Biol Chem. 1970 Oct 25;245(20):5382–5389. [PubMed] [Google Scholar]
  33. Teng C. S., Hamilton T. H. Regulation by estrogen of organ-specific synthesis of a nuclear acidic protein. Biochem Biophys Res Commun. 1970 Sep 10;40(5):1231–1238. doi: 10.1016/0006-291x(70)90927-7. [DOI] [PubMed] [Google Scholar]
  34. Teng C. T., Teng C. S., Allfrey V. G. Species-specific interactions between nuclear phosphoproteins and DNA. Biochem Biophys Res Commun. 1970 Nov 9;41(3):690–696. doi: 10.1016/0006-291x(70)90068-9. [DOI] [PubMed] [Google Scholar]
  35. Tveter K. J., Attramadal A. Selective uptake of radioactivity in rat ventral prostate following administration of testosterone-1,2-3H. Methodological considerations. Acta Endocrinol (Copenh) 1968 Oct;59(2):218–226. doi: 10.1530/acta.0.0590218. [DOI] [PubMed] [Google Scholar]
  36. Widnell C. C., Tata J. R. A procedure for the isolation of enzymically active rat-liver nuclei. Biochem J. 1964 Aug;92(2):313–317. doi: 10.1042/bj0920313. [DOI] [PMC free article] [PubMed] [Google Scholar]

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