Acceptor proteins in rat androgenic tissue chromatin.

Klyzsejko-Stefanowicz, Leokadia; Chiu, Jen‐Fu; Tsai, Yu‐Hui; Hnilica, Lubomir S.

doi:10.1073/pnas.73.6.1954

Cited by 55 publications

(24 citation statements)

References 28 publications

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“…However, it may be the lengths and sequences of the (A+T)-rich regions that are important, not just the A+T composition of the genome. These data and previous data from our and other laboratories (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) appear to conflict with another model of acceptor sites wherein specific DNA sequences are recognized directly by steroid receptors (44)(45)(46)(47)(48). The existence of two classes of sites, one containing both DNA and protein and one containing just DNA, has been suggested (49)(50)(51).…”

Section: Discussionmentioning

confidence: 31%

“…Nuclear acceptor sites for the avian oviduct estrogen receptor have also been reported to be composed ofDNA and a tightly bound protein in the CP-3 fraction (12), although these sites appear to be distinct from the acceptor sites for PR (13). Similarly, a role for tightly bound proteins and DNA in the nuclear acceptor sites for androgens, estrogens, progesterones, and glucocorticoids in mammalian tissues has been reported (12,(14)(15)(16)(17). It has recently been shown that the number of PR binding sites on hen DNA, generated by rehybridizing increasing quantities of acceptor protein to the hen DNA, is limited (6), suggesting that specific DNA sequences may be involved in the nuclear acceptor sites for PR.…”

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confidence: 99%

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Evidence for specific DNA sequences in the nuclear acceptor sites of the avian oviduct progesterone receptor.

Toyoda¹,

Seelke²,

Littlefield³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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Nuclear acceptor sites (binding sites) for the avian oviduct progesterone receptor (PR) have been reported to involve both DNA and specific chromatin proteins in the tightly bound fraction termed CP-3 (1-6). These acceptor sites bind the PR with high affinity and saturability. Similarities between cell-free nuclear binding to these sites using isolated PR and nuclear binding in vivo have been reported (2, 3, 7-11). Nuclear acceptor sites for the avian oviduct estrogen receptor have also been reported to be composed ofDNA and a tightly bound protein in the CP-3 fraction (12), although these sites appear to be distinct from the acceptor sites for PR (13). Similarly, a role for tightly bound proteins and DNA in the nuclear acceptor sites for androgens, estrogens, progesterones, and glucocorticoids in mammalian tissues has been reported (12,(14)(15)(16)(17). It has recently been shown that the number of PR binding sites on hen DNA, generated by rehybridizing increasing quantities of acceptor protein to the hen DNA, is limited (6), suggesting that specific DNA sequences may be involved in the nuclear acceptor sites for PR. In this paper, we attempt to substantiate this possibility by determining whether or not DNA from other species can bind to the hen oviduct acceptor protein to generate nuclear acceptor sites for the PR. MATERIALS AND METHODSIsolation and Binding of the PR and Nuclear Components. The methods and materials used in these studies have been described (6). The PR binding assay used the streptomycin assay (18) with modification in the measurement of radioactivity. The method ofisolating the nonfunctional PR, which can bind the steroid but cannot bind the nuclear acceptor sites in vivo or in vitro, is described elsewhere (7,8,10).Nuclei and chromatin were isolated and purified from these homogenates by using modifications of previously described methods (7). All steps were performed at 0-40C. Isolation of the native nucleoacidic protein (NAP) and pure hen DNA has been described (4, 6, 18). Chromatin, NAP, and DNA were suspended in 4 mM Tris.HCl/0.2 mM EDTA, pH 7.5, at 0.5-1.0 mg ofDNA/ml for use in the PR binding assays. CP-3 protein was isolated from hen oviduct chromatin by using hen oviduct chromatin-hydroxylapatite chromatography as described (6). This chromatin-hydroxylapatite resin was treated with a stepwise gradient of increasing concentrations of guanidinium hydrochloride (Gdn.HCl)-0, 4, and 7 M-in 0.1 M sodium phosphate buffer, pH 6.0, at 40C. The ratio of solvent to resin (ml/g of resin) was -5 with a flow rate of 5.0 ml/min. The protein concentration in the eluting fractions was determined by the method of Bramhall et al. (19) substituting Coomassie blue stain or by the method of Bradford (20). In later studies, the method of Bradford was used for protein quantitation.Reconstitution of CP-3 to DNA to Obtain Reconstituted NAP Containing PR Binding Sites. The method for reconstituting the NAP that results in optimal recoveries of acceptor sites for PR when using the hen oviduct chromos...

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Section: Discussionmentioning

confidence: 31%

mentioning

confidence: 99%

Evidence for specific DNA sequences in the nuclear acceptor sites of the avian oviduct progesterone receptor.

Toyoda¹,

Seelke²,

Littlefield³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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“…In many different target-tissue systems a significant amount of steroid-binding activity remains resistant to this solubilization (see Barrack & Coffey, 1982, and references quoted therein). Significant roles have been proposed for the salt-resistant sites (Clark & Peck, 1976;Ruh & Baudendistel, 1978) and specific acceptor activity has been implicated (Davies & Griffiths, 1973;Klyzsejko-Stefanowicz, Chiù, Tsai & Hnilica, 1976;Wang, 1978). Objections to their physiological significance have been offset to some degree by arguments relating to dose-responses and methodology of labelling or assessment (Barrack, Hawkins, Allen, Hicks & Coffey, 1977; Barrack, Hawkins & Coffey, 1979;Barrack & Coffey, 1980.…”

Section: Introductionmentioning

confidence: 99%

Extraction of androgen–receptor complexes from regions of rat ventral prostate nuclei sensitive or resistant to nucleases

Davies¹

1983

Journal of Endocrinology

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Rat ventral prostate nuclei contain androgen-binding sites which are susceptible or resistant to excision by endonucleolytic action. Those which were susceptible were associated both with oligonucleosomal and subnucleosomal particles. The sedimentation profile characteristic of a nuclear androgen-receptor complex could be obtained by exhaustive nucleolytic digestion or by treatment of fractions with KCl (0.6 mol/l). Androgen-binding sites resistant to DNAase I were also resistant to KCl, whereas those sites resistant to micrococcal nuclease were partially extractable with KCl. Nuclease-resistant sites could be extracted with heparin (10 mg/ml). Androgen-receptor complexes obtained from nuclease-sensitive or nuclease-resistant regions by extraction with KCl or heparin were indistinguishable by routine sedimentation analysis.

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“…In other organisms, similar nuclear acceptor sites composed of tightly bound (to DNA) nonhistone proteins have been reported for the estrogen receptor/chicken oviduct system (17), for systems involving the estrogen and progesterone receptors in sheep brain (22), in hamster uteri (t), in cow, rabbit, and human uteri (21,23,24), for systems involving the glucocorticoid receptor in both rat liver (25) and human leukemic cell line system (26), and finally for the androgen receptor rat prostate system (27). Thus, the acceptor protein-DNA complex as a potential nuclear acceptor site model for steroid receptors seems to be universal.…”

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confidence: 99%

Nuclease resistance and the enrichment of native nuclear acceptor sites for the avian oviduct progesterone receptor.

Hora¹,

Horton²,

Toft³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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High-affinity nucleoprotein acceptor sites for the avian oviduct progesterone receptor (PR) have been enriched by a combination of nuclease digestion and centrifugation. These enriched binding elements exhibited markedly enhanced PR binding on a per mass DNA basis compared to chromatin (20-to 25-fold) or dehistonized chromatin (4-to 5-fold). Electrophoretic analysis of the nuclease-resistant DNA showed that there is a set of DNA fragments of 100-150 base pairs that are protected from digestion. Excessive digestion resulted in smaller DNA fragments and a loss of PR binding activity. The PR binding was saturable using a crude receptor preparation and displayed a competition with the same receptor preparation that was labeled with nonradioactive progesterone. The enhanced binding was also demonstrable using highly purified receptor preparations that exhibit two classes of binding sites both of which are of high affinity and saturable as assessed by Scatchard analyses. These two high-affinity classes of binding sites are shown to be competed by unlabeled purified PR. The nuclease resistance of these nucleoprotein acceptor sites from chromatin is a property similar to the nuclear matrix binding sites suggesting a relationship between these two classes of nuclear acceptor sites.Based on the presently accepted mechanism of steroid hormone action, the interaction of a steroid receptor-hormone complex with the nucleus is an important step in the induction of specific gene products. A great deal of effort has been invested by a number of laboratories in trying to determine the relevant receptor-nuclear interactions. Despite these efforts, the exact nature of the nuclear binding sites (acceptor sites) is not completely understood. Steroid receptors have been shown to bind to DNA (1), RNA (2), chromatin (3), and ribonucleoprotein particles (4).This laboratory has been concentrating on the specific interaction ofthe chicken oviduct progesterone receptor (PR) with nuclear acceptor sites in oviduct chromatin. In contrast to the receptor binding to pure DNA sequences alone (5, 6), the binding of the oviduct PR to subfractions of chromatin containing nonhistone protein-DNA complexes, termed nucleoacidic protein or NAP, has been shown to be receptor dependent and saturable, of high affinity (7)(8)(9)(10)(11), to be receptor specific (12), and to generate patterns of bindings similar to that measured in vivo (13)(14)(15)(16). Further, the capacity of the PR to bind to nuclear acceptor sites reflects the ability of progesterone to alter RNA polymerase activity in nuclear run-off experiments (17, t) and to specifically induce the avidin gene (18). A specific subset of nonhistone proteins (acceptor proteins) has been shown to be necessary for the generation of specific nucleoprotein acceptor sites (10,11,(19)(20)(21).In other organisms, similar nuclear acceptor sites composed of tightly bound (to DNA) nonhistone proteins have been reported for the estrogen receptor/chicken oviduct system (17), for systems involving the estrog...

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Acceptor proteins in rat androgenic tissue chromatin.

Cited by 55 publications

References 28 publications

Evidence for specific DNA sequences in the nuclear acceptor sites of the avian oviduct progesterone receptor.

Evidence for specific DNA sequences in the nuclear acceptor sites of the avian oviduct progesterone receptor.

Extraction of androgen–receptor complexes from regions of rat ventral prostate nuclei sensitive or resistant to nucleases

Nuclease resistance and the enrichment of native nuclear acceptor sites for the avian oviduct progesterone receptor.

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