Purification of a nuclear protein (Receptor Binding Factor-1) associated with the chromatin acceptor sites for the avian oviduct progesterone receptor

Rejman, J. J.; Landers, James P.; Goldberger, Amy; McCormick, Daniel J.; Gosse, Barbara; Spelsberg, Thomas C.

doi:10.1007/bf01025717

Cited by 8 publications

(14 citation statements)

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“…The RBF-1 protein appears to play a role in the highest affinity class of nuclear binding sites for PR in the avian oviduct [Rejman et al, 1991;Schuchard et al, 1991a;Horton et al, 19911. Selective removal of the chromatin protein fraction which contains RBF-1 results in the loss of the highest affinity class of PR binding sites.…”

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confidence: 97%

Immunohistochemical localization of the avian progesterone receptor and its candidate receptor binding factor (RBF‐1)

Zhuang

Landers

Schuchard

et al. 1993

J of Cellular Biochemistry

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An avian oviduct nuclear matrix protein in the 6-10 kDa size range has been implicated to function in the cell-free nuclear binding of the avian oviduct progesterone receptor (PR). This protein, termed the receptor binding factor-1 (RBF-1), has been purified and partially characterized [Schuchard et al.: Biochemistry 30:4535-4542, 1991]. This paper describes the immunohistochemical co-localization of the RBF-1 and PR in the avian oviduct cell nuclei and rat reproductive cell nuclei using antibodies directed specifically against the RBF-1 and activated PR. In the undifferentiated oviduct, the immunoreactivities for both PR and RBF-1 were co-localized in the nuclei of only epithelial cells, but not the stromal cells or smooth muscle cells. In the partially differentiated oviduct of estrogen treated chicks, the immunoreactivity co-localized in the nuclei of not only epithelial but also glandular and stromal cells. Staining for the PR, but not RBF-1, was detected in the smooth muscle cells. The intensity of the PR but not the RBF-1 staining was markedly down-regulated in these cells at 2 and 6 h after treatment of the animals with progesterone (P). However, the band patterns for RBF-1 in the Western blots did show qualitative changes which may reflect P-induced posttranslational modifications which alter the epitope on the RBF-1. Interestingly, immunohistochemical analysis of several reproductive tissues of the rat showed that certain cell types in the uterus, ovary, and prostate displayed strong positive nuclear staining for an RBF-1-like antigen(s).(ABSTRACT TRUNCATED AT 250 WORDS)

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confidence: 97%

Immunohistochemical localization of the avian progesterone receptor and its candidate receptor binding factor (RBF‐1)

Zhuang

Landers

Schuchard

et al. 1993

J of Cellular Biochemistry

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“…Briefly, purified RBF was solubilized in 6 M guanidine-HCl in reconstitution buffer (10 mM Tris, 1 mM EDTA, 1 mM dithiothreitol, 0.2 mM PMSF) to a concentration of 250 ng/ml. This concentration of the RBF was found to be important for optimal protein renaturation and reconstitution of PR-binding sites on the DNA (26,30,48). It was also found in these current studies to be important for achieving the optimal DNA sequence-specific binding.…”

Section: Isolation and Purification Of Rbf By Preparativementioning

confidence: 56%

“…In our laboratory, a specific "acceptor protein," RBF, was isolated based on its ability to generate saturable, high affinity binding sites on avian and mammalian genomic DNAs and characterized as a component of the chromatin nuclear matrix acceptor sites for the avian oviduct PR (22,23,25,26,29,30,67,68). The RBF failed to generate these sites when using insect and prokaryotic genomic DNA.…”

Section: Discussionmentioning

confidence: 99%

“…Reconstitution of Purified RBF-This method has been described in detail previously (22,26,48). Briefly, purified RBF was solubilized in 6 M guanidine-HCl in reconstitution buffer (10 mM Tris, 1 mM EDTA, 1 mM dithiothreitol, 0.2 mM PMSF) to a concentration of 250 ng/ml.…”

Section: Isolation and Purification Of Rbf By Preparativementioning

confidence: 99%

“…Previous studies in our laboratory revealed that chromatin acceptor sites for the avian oviduct progesterone receptor (PR) were associated with the nuclear matrix as was the receptor-binding factor (RBF) (23)(24)(25). In short, the chromatin and nuclear matrix acceptor sites appear to be one and the same.We have purified the nuclear matrix RBF (formerly termed a chromatin "acceptor" protein) based on its ability to generate specific, high affinity PR binding when renatured and bound to the avian genomic DNA (12,22,23,26). Using immunohistochemical techniques with antibodies against the purified RBF, the RBF was localized to the nuclei of selected cell types in certain avian and rat tissues (27,28).…”

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A DNA-binding Element for a Steroid Receptor-binding Factor Is Flanked by Dual Nuclear Matrix DNA Attachment Sites in the c-myc Gene Promoter

Lauber

Barrett

Subramaniam

et al. 1997

Journal of Biological Chemistry

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The receptor-binding factor (RBF) for the avian oviduct progesterone (Pg) receptor (PR) has previously been shown to be a unique 10-kDa nuclear matrix protein that generates high affinity PR-binding sites on avian DNA. This paper describes the use of Southwestern blot and DNA gel shift analyses with RBF protein to identify a minimal 54-base pair RBF-binding element in the matrix-associated region (MAR) of the Pg-regulated c-myc gene promoter. This element contains a 5 -GC-rich domain and a 3 -AT-rich domain, the latter of which has a homopurine/homopyrimidine structure. The gel shift assays required the generation of an RBF-maltose fusion protein (RBF-MBP), which specifically binds this element and is supershifted when the anti-RBF polyclonal antibody is added. Computer analysis of the fulllength amino acid sequence for RBF predicts a DNAbinding motif involving a ␤-sheet structure at the N-terminal domain. Southern blot analyses using nuclear matrix DNA suggests that there are dual MAR sites in the c-myc promoter, which flank an intervening domain containing the RBF element. The co-transfection of this MAR sequence, containing the RBF element and cloned into a luciferase reporter vector, together with an RBF expression vector construct, into steroid treated human MCF-7 cells, results in a decrease of the c-myc promoter activity relative to control transfections containing only the parent vector of the RBF expression construct. These data suggest that a unique chromatin/ nuclear matrix structure, composed of the RBF-DNA element complex which is flanked by nuclear matrix attachment sites, serves to bind the PR and repress the c-myc promoter.The classic model of steroid hormone action suggests that ligand-bound receptors dimerize and bind to specific sites (acceptor sites) on the chromatin/DNA to alter gene expression. In many genes, the steroid receptors (SR) 1 bind to specific cisacting DNA sequences, referred to as hormone response elements (for reviews, see Refs. 1-3). These elements function as enhancer-like DNA sequences and are required for the steroid hormone regulation of transcription. However, not all steroidresponsive genes contain these elements, and several other pathways and participating nuclear proteins have now been identified. The SR regulation of gene transcription has now been shown to involve 1) interactions of the SR with transcription factors, such as the AP-1 complex (for review, see Refs. 2-4); 2) interactions of SR with accessory factors, which appear to function by assisting the binding of the receptor to the steroid response elements, or to transcription factors (for reviews see Refs. 4 -6 and references therein); and finally, 3) interactions of SR with specific chromatin structures including the nuclear matrix (see Refs. 7-15 and references therein). In the latter, a class of high affinity binding sites (acceptor sites) for SRs in the nuclei, chromatin, and nuclear matrix DNAbinding proteins (e.g. acceptor proteins) has been reported by many laboratories for most of the steroid hormone...

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The ubiquitous nature of the progesterone receptor binding factor‐1 (RBF‐1) in avian tissues

Landers

Subramaniam

Gosse

et al. 1994

J of Cellular Biochemistry

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The avian oviduct receptor binding factor-1 (RBF-1) is a 10 kDa nuclear matrix protein that was originally identified through its ability to effect high affinity interaction of activated progesterone receptor (PR) with chromatin. In the present study, the RBF-1 is shown to not be restricted to reproductive tissues (e.g., oviduct) but present in all avian tissues examined by Western blot analysis with a monoclonal antibody prepared against purified RBF-1. The heart and pancreas had the highest and lowest RBF-1 levels, respectively; the concentration ranging by approximately 50-fold in these tissues. The 10 kDa size of the RBF-1 detected in all tissues suggests no significant tissue-specific differences in the protein. This was consistent with the finding that purified hepatic and oviductal RBF-1 have identical amino-terminal sequence. Using a recently isolated cDNA to RBF-1, the levels of RBF-1 mRNA were found to correlate well with the ubiquitous presence of the protein as well as tissue-specific differences in concentration. The presence of RBF-1 in non-progesterone responsive tissues suggests the possibility that RBF-1 may not be specifically involved in PR-DNA interactions but may play a more diverse role, possibly involving other steroid receptors such as the glucocorticoid receptor.

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Purification of a nuclear protein (Receptor Binding Factor-1) associated with the chromatin acceptor sites for the avian oviduct progesterone receptor

Cited by 8 publications

References 71 publications

Immunohistochemical localization of the avian progesterone receptor and its candidate receptor binding factor (RBF‐1)

Immunohistochemical localization of the avian progesterone receptor and its candidate receptor binding factor (RBF‐1)

A DNA-binding Element for a Steroid Receptor-binding Factor Is Flanked by Dual Nuclear Matrix DNA Attachment Sites in the c-myc Gene Promoter

The ubiquitous nature of the progesterone receptor binding factor‐1 (RBF‐1) in avian tissues

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