Characterization of a purified chromatin acceptor protein (receptor binding factor 1) for the avian oviduct progesterone receptor

M. Schuchard, J. J. Rejman, Daniel J Mc Cormick, B. Gosse, T. Ruesink, T. C. Spelsberg

Research output: Contribution to journalArticle

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Abstract

The specific, high-affinity binding of the avian oviduct progesterone receptor (PR) with target-cell nuclei and chromatin has been shown to involve DNA complexed with specific chromatin acceptor proteins. One of these chromatin acceptor proteins has been partially purified and found to be a small hydrophobic protein with a broad pI of 5.0-6.0 [Goldberger, A., and Spelsberg, T.C., (1988) Biochemistry 27, 2103-2109]. This paper describes the final purification over 100,000-fold to apparent homogenity of this candidate PR acceptor protein, termed the receptor binding factor 1 (RBF-1). When the avian genomic DNA is bound by RBF-1, saturable, high-affinity (K(D) ~ 2 x 10-9 M) binding sites for PR are generated. RBF-1 has a unique, hydrophobic N-terminal sequence. The PR binding to the RBF-1-DNA complexes is shown to be dependent on an intact activated PR with which excess nonradiolabeled PR can compete. By use of a new, highly specific monoclonal antibody (mAb) to the RBF-1 with Western immunoblotting, RBF-1 was shown to be localized in the nucleus and to be tissue and species specific. Selective removal of the chromatin proteins containing RBF-1 results in the loss of the highest affinity class of PR binding sites. A second class of residual PR binding sites remains in the nucleoacidic protein (NAP), a complex of proteins more tightly bound to the DNA. This class of PR binding activity has been classified as the RBF-2. The RBF-1 is estimated to be 0.03% of the total chromatin protein with about 1.2 x 105 molecules/diploid cell.

Original languageEnglish (US)
Pages (from-to)4535-4542
Number of pages8
JournalBiochemistry
Volume30
Issue number18
StatePublished - 1991

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Oviducts
Progesterone Receptors
Protein Binding
Chromatin
Proteins
DNA
Binding Sites
Biochemistry
Cell Nucleus
Diploidy
Purification
Western Blotting
Monoclonal Antibodies
Cells
Tissue
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of a purified chromatin acceptor protein (receptor binding factor 1) for the avian oviduct progesterone receptor. / Schuchard, M.; Rejman, J. J.; Mc Cormick, Daniel J; Gosse, B.; Ruesink, T.; Spelsberg, T. C.

In: Biochemistry, Vol. 30, No. 18, 1991, p. 4535-4542.

Research output: Contribution to journalArticle

Schuchard, M, Rejman, JJ, Mc Cormick, DJ, Gosse, B, Ruesink, T & Spelsberg, TC 1991, 'Characterization of a purified chromatin acceptor protein (receptor binding factor 1) for the avian oviduct progesterone receptor', Biochemistry, vol. 30, no. 18, pp. 4535-4542.
Schuchard, M. ; Rejman, J. J. ; Mc Cormick, Daniel J ; Gosse, B. ; Ruesink, T. ; Spelsberg, T. C. / Characterization of a purified chromatin acceptor protein (receptor binding factor 1) for the avian oviduct progesterone receptor. In: Biochemistry. 1991 ; Vol. 30, No. 18. pp. 4535-4542.
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