Binding of the glucocorticoid receptor to the rat liver nuclear matrix. The role of disulfide bond formation

Scott H Kaufmann, S. Okret, A. C. Wikstrom, J. A. Gustafsson, J. H. Shaper

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

The nuclear matrix is a putative skeletal structure which has been implicated in many nuclear functions. To assess a possible role of the nuclear matrix in glucocorticoid action, purified rat liver nuclei containing glucocorticoid-receptor complexes were treated with DNase I ± RNase A followed by 1.6 M NaCl, thus yielding salt-extractable and salt-resistant (nuclear matrix) fractions. The subnuclear distribution of hormone-receptor complexes was determined by (i) following the fate of unmetabolized radiolabel after injection of labeled triamicinolone acetonide into adrenalectomized animals and (ii) subjecting various subfractions to immunoblotting using a monoclonal antibody which recognizes the glucocorticoid receptor. Both techniques indicated that 50-70% of the total nuclear hormone-receptor complexes were recovered in the nuclear matrix fraction. Previous results (Kaufmann, S.H., and Shaper, J.H. (1984) Exp. Cell Res. 155, 477-495) suggest that a variety of nuclear polypeptides become nuclease- and salt-resistant as a result of the formation of intermolecular disulfide bonds. The following evidence suggests that disulfide bonds mediate the association between the glucocorticoid receptor and the nuclear matrix. (i) When nuclei were isolated in the absence of sulfhydryl-blocking and -cross-linking reagents, sodium dodecyl sulfate-polyacryl-amide gel electrophoresis under nonreducing conditions revealed that the receptor was present as a high molecular weight disulfide-cross-linked complex. (ii) When nuclei were isolated in the presence of the irreversible sulfhydryl-blocking reagent iodoacetamide, the disulfide bonds which cross-linked the receptor into high molecular weight complexes were absent; and 85-100% of the hormone-receptor complexes were salt-extractable. (iii) When nuclei (isolated in the absence of iodoacetamide) were treated with the sulfhydryl-cross-linking reagent sodium tetrathionate, >95% of the nuclear hormone-receptor complexes became resistant to extraction with nucleases and 1.6 M NaCl. The implications of these results for other matrix-associated nuclear functions are discussed.

Original languageEnglish (US)
Pages (from-to)11962-11967
Number of pages6
JournalJournal of Biological Chemistry
Volume261
Issue number26
StatePublished - 1986
Externally publishedYes

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Nuclear Matrix
Glucocorticoid Receptors
Disulfides
Liver
Rats
Salts
Cross-Linking Reagents
Iodoacetamide
Cytoplasmic and Nuclear Receptors
Tetrathionic Acid
Molecular Weight
Molecular weight
Hormones
Sulfhydryl Reagents
Pancreatic Ribonuclease
Deoxyribonuclease I
Electrophoresis
Immunoblotting
Amides
Sodium Dodecyl Sulfate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding of the glucocorticoid receptor to the rat liver nuclear matrix. The role of disulfide bond formation. / Kaufmann, Scott H; Okret, S.; Wikstrom, A. C.; Gustafsson, J. A.; Shaper, J. H.

In: Journal of Biological Chemistry, Vol. 261, No. 26, 1986, p. 11962-11967.

Research output: Contribution to journalArticle

Kaufmann, Scott H ; Okret, S. ; Wikstrom, A. C. ; Gustafsson, J. A. ; Shaper, J. H. / Binding of the glucocorticoid receptor to the rat liver nuclear matrix. The role of disulfide bond formation. In: Journal of Biological Chemistry. 1986 ; Vol. 261, No. 26. pp. 11962-11967.
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