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PMID:2021645

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Citation

Schuchard, M, Rejman, JJ, McCormick, DJ, Gosse, B, Ruesink, T and Spelsberg, TC (1991) Characterization of a purified chromatin acceptor protein (receptor binding factor 1) for the avian oviduct progesterone receptor. Biochemistry 30:4535-42

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., & Spelsberg, T. C., (1988) Biochemistry 27, 2103-2109]. This paper describes the final purification over 100,000-fold to apparent homogeneity 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 (KD approximately 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 10(5) molecules/diploid cell.

Links

PubMed

Keywords

Amino Acid Sequence; Animals; Avian Proteins; Binding, Competitive; Blotting, Western; Carrier Proteins/genetics; Carrier Proteins/immunology; Carrier Proteins/isolation & purification; Chickens; Chromatin/chemistry; DNA/metabolism; DNA-Binding Proteins/genetics; DNA-Binding Proteins/immunology; DNA-Binding Proteins/isolation & purification; Molecular Sequence Data; Oviducts/metabolism; Receptors, Progesterone/genetics; Receptors, Progesterone/immunology; Receptors, Progesterone/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

CHICK:PRGR

located_in

GO:0000785: chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(UBERON:0000993)
  • part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:Q7LZ14

located_in

GO:0000785: chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(UBERON:0000993)
  • part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:Q9YH14

located_in

GO:0000785: chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(UBERON:0000993)|part_of:(UBERON:0002106)|part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:PRGR

part_of

GO:0032993: protein-DNA complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:PRGR

part_of

GO:0000790: nuclear chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(UBERON:0000993)
  • part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:PRGR

enables

GO:0031490: chromatin DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:Q7LZ14

part_of

GO:0032993: protein-DNA complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:Q7LZ14

enables

GO:0031490: chromatin DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:Q7LZ14

part_of

GO:0000790: nuclear chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(UBERON:0000993)
  • part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:Q7LZ14

enables

GO:0033142: progesterone receptor binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:Q9YH14

part_of

GO:0000790: nuclear chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(UBERON:0000993)|part_of:(UBERON:0002106)|part_of:(UBERON:0002107)

Seeded From UniProt

complete

CHICK:Q9YH14

enables

GO:0033142: progesterone receptor binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CHICK:Q9YH14

part_of

GO:0032993: protein-DNA complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHICK:Q9YH14

enables

GO:0031490: chromatin DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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