Proteins that mask the nuclear binding sites of the avian oviduct progesterone receptor

Biochemistry ◽  
1985 ◽  
Vol 24 (24) ◽  
pp. 6988-6997 ◽  
Author(s):  
Ginger Martin Dani ◽  
Thomas C. Spelsberg
Keyword(s):  
Progesterone Receptor ◽  
Binding Sites ◽  
Nuclear Binding

scholarly journals Nuclear binding of progesterone in hen oviduct. Role of acidic chromatin proteins in high-affinity binding

1976 ◽  
Vol 156 (2) ◽  
pp. 409-418 ◽  
Author(s):  
R A Webster ◽  
G M Pikler ◽  
T C Spelsberg
Keyword(s):  
Progesterone Receptor ◽  
Binding Sites ◽  
Acidic Protein ◽  
Affinity Binding ◽  
Target Tissue ◽  
High Affinity Binding ◽  
Receptor Complexes ◽  
High Affinity ◽  
Nuclear Binding ◽  
Acidic Proteins

The multiple classes of binding sites for the progesterone-receptor complex in hen oviduct muclei were found to be of chromatin origin. The highest-affinity, and presumably most physiologically important class, is localized in oviduct chromatin and contains approx. 6000-10000 sites per nucleus. None of these sites is detected in spleen chromatin. Two new techniques were used for assaying rapidly the binding of steroid-receptor complexes to soluble deoxyribonucleoproteins in vito. The extent of high-affinity binding by the nucleo-acidic protein fraction from spleen chromatin is as great as that by the nucleo-acidic protein from oviduct chromatin. Consequently the tissue-specific nuclear binding of the progesterone receptor is found not to be a consequence of the absence of the nuclear binding sites (acceptors) from chromatin of non-target tissue (spleen), but rather a result of complete masking of these sites. In the target-tissue (oviduct) chromatin, approx. 70% of the high-affinity acceptor sites are also masked. Acidic proteins, and not histones, appear to be responsible for the masking of these acceptor sites. In addition, acidic proteins represent (or at least are an essential part of) these high-affinity sites in the oviduct nucleus. Pure DNA displays a few high-and many low-affinity binding sites. In support of previous work with immature chicks, the acidic protein fraction of the nucleo-acidic results thus support the hypotheis that protein complexed with DNA, and not DNA alone, represent the high-affinity binding sites for the steroid-receptor complexes in nuclear chromatin. The lower-affinity classes of binding sites may represent DNA and/or other nuclear components.

79 Specific proteins and DNA sequences as nuclear binding sites for the avian oviduct progesterone receptor

1983 ◽  
Vol 19 ◽  
pp. 27
Author(s):  
Thomas C. Spelsberg ◽  
Ralph Seelke ◽  
Bruce Littlefield ◽  
Hiroo Toyoda ◽  
Thoas Ruh
Keyword(s):  
Progesterone Receptor ◽  
Dna Sequences ◽  
Binding Sites ◽  
Nuclear Binding ◽  
Specific Proteins

Monoclonal Antibodies Recognizing the Nuclear Binding Sites of the Avian Oviduct Progesterone Receptor*

Endocrinology ◽  
1986 ◽  
Vol 118 (6) ◽  
pp. 2235-2241 ◽  
Author(s):  
AMY GOLDBERGER ◽  
BRUCE A. LITTLEFIELD ◽  
JERRY KATZMANN ◽  
THOMAS C. SPELSBERG
Keyword(s):  
Monoclonal Antibodies ◽  
Progesterone Receptor ◽  
Binding Sites ◽  
Nuclear Binding

scholarly journals Nuclear binding of progesterone in hen oviduct. Binding to multiple sites in vitro

1976 ◽  
Vol 156 (2) ◽  
pp. 399-408 ◽  
Author(s):  
G M Pikler ◽  
R A Webster ◽  
T C Spelsberg
Keyword(s):  
Progesterone Receptor ◽  
Binding Site ◽  
Cell Nucleus ◽  
Binding Sites ◽  
Nuclear Material ◽  
Target Cells ◽  
Target Tissue ◽  
Nuclear Binding

Steroid hormones, including progesterone, are known to bind with high affinity (Kd approximately 1 × 10(-10)M) to receptor proteins once they enter target cells. This complex (the progesterone-receptor) then undergoes a temperature-and/or salt-dependent activation which allows it to migrate to the cell nucleus and to bind to the deoxyribonucleoproteins. The present studies demonstrate that binding the hormone-receptor complex in vitro to isolated nuclei from the oviducts of laying hens required the same conditions as do other studies of bbinding in vitro reported previously, e.g. the hormone must be complexed to intact and activated receptor. The assay of the nuclear binding by using multiple concentrations of progesterone receptor reveals the presence of more than one class of binding site in the oviduct nuclei. The affinity of each of these classes of binding sites range from Kd approximately 1×10(-9)-1×10(-8)M. Assays using free steroid (not complexed with receptor) show no binding to these sites. The binding to each of the classes of sites, displays a differential stability to increasing ionic concentrations, suggesting primarily an ionic-type interaction for all classes. Only the highest-affinity class of binding site is capable of binding progesterone receptor under physioligical-saline conditions. This class represent 6000-10000 sites per cell nucleus and resembles the sites detected in vivo (Spelsberg, 1976, Biochem. J. 156, 391-398) which cause maximal transcriptional response when saturated with the progesterone receptor. The multiple binding sites for the progesterone receptor either are not present or are found in limited numbers in the nuclei of non-target organs. Differences in extent of binding to the nuclear material between a target tissue (oviduct) and other tissues (spleen or erythrocyte) are markedly dependent on the ionic conditions, and are probably due to binding to different classes of sites in the nuclei.

Estrogen and progesterone receptor-binding sites on the chicken vitellogenin II gene: synergism of steroid hormone action

1988 ◽  
Vol 8 (12) ◽  
pp. 5323-5330
Author(s):  
A C Cato ◽  
E Heitlinger ◽  
H Ponta ◽  
L Klein-Hitpass ◽  
G U Ryffel ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Receptor Binding ◽  
Dna Sequences ◽  
Binding Sites ◽  
Consensus Sequence ◽  
Chloramphenicol Acetyltransferase ◽  
Synergistic Action ◽  
Gene Fragment ◽  
T47d Cells ◽  
Vitellogenin Gene

The chicken vitellogenin II gene is transcriptionally activated by estrogens. In transient transfection experiments in human T47D cells that contain receptors for various steroids, we showed estradiol, progestin, and androgen responses of a chimeric chicken vitellogenin II construct. This construct consists of DNA sequences from -626 to -590 upstream of the start of transcription of the chicken vitellogenin gene linked to the herpes simplex virus thymidine kinase promoter driving the transcription of the bacterial chloramphenicol acetyltransferase gene. Treatment of the transfected T47D cells with a combination of estradiol and the progestin R5020 led to a superinduction of chloramphenicol acetyltransferase activity, showing a synergistic action of these two steroids. This synergism was not observed upon treatment of the transfected cells with estradiol and the androgen dihydrotestosterone. Using point mutations in the vitellogenin gene fragment, we showed in functional and in in vitro DNase I footprinting assays with a purified progesterone receptor that, for the synergistic action of estradiol and R5020 to occur, the progesterone receptor must be bound to the vitellogenin gene fragment. The progesterone receptor-binding site was localized at -610 to -590, close to the consensus sequence (-626 to -613) for estrogen receptor binding and function. We therefore demonstrate here that two different steroid hormones can be functionally synergistic through the interaction of their corresponding receptors with two different binding sites adjacent to one another.

Characterization of the Nuclear Binding Sites (Acceptor Sites) for a Steroid Receptor

1987 ◽  
pp. 111-136 ◽  
Author(s):  
T. Spelsberg ◽  
A. Goldberger ◽  
J. Hora ◽  
M. Horton ◽  
B. Littlefield
Keyword(s):  
Binding Sites ◽  
Steroid Receptor ◽  
Nuclear Binding

Corticosterone's metabolite is an agonist for Na+ transport stimulation in A6 cells

1988 ◽  
Vol 255 (4) ◽  
pp. F736-F748 ◽  
Author(s):  
R. L. Duncan ◽  
W. M. Grogan ◽  
L. B. Kramer ◽  
C. O. Watlington
Keyword(s):  
Binding Sites ◽  
Short Circuit ◽  
Type I ◽  
Type Ii ◽  
Na Transport ◽  
Single Class ◽  
Apparent Dissociation Constant ◽  
Lower Affinity ◽  
A6 Cells ◽  
Nuclear Binding

This study tests the hypothesis, in A6 epithelia, that 1) corticosterone stimulates active Na+ transport (short-circuit current, Isc) by an additional receptor mechanism to the type I (mineralocorticoid) and type II (glucocorticoid) mechanisms shared with aldosterone (Aldo) and 2) that the agonist may be 6 beta-OH-corticosterone made in the effector cell. The dose-response relationship of corticosterone at 24 h resolves into two components, by curve fitting, with a 50% effective concentration (EC50) for 10% of maximum Isc stimulation of 2 X 10(-9) M and an EC50 for the other 90% of 3 X 10(-7) M. The EC50 of the smaller component correlates with the apparent dissociation constant (K'd) of corticosterone for high affinity (type II) nuclear binding sites shared with Aldo. In unlabeled analogue competition studies Aldo and corticosterone displaced nuclear binding equally below 10(-8) M [3H]corticosterone, indicating only shared sites. However, nonshared saturable sites (displaced by corticosterone but not by Aldo) were found at [3H]-corticosterone concentrations above 10(-8) M. Concentration-binding curves performed with [3H]corticosterone, in presence of 1,000 X Aldo to displace shared sites, revealed a single class of binding sites with a half-maximal saturation of 2 X 10(-7) M, which is quite similar to the EC50 of the lower affinity component of Isc stimulation by corticosterone at 24 h. Reversed phase high-pressure liquid chromatography of nuclear extracts indicates that the saturable component of bound [3H] was 6 beta-OH-[3H]corticosterone derived from [3H]corticosterone. Thus, A6 cells metabolize corticosterone to 6 beta-OH-corticosterone, which in turn occupies lower-affinity receptors not shared with Aldo or corticosterone, to mediate most of the active Na+ transport stimulation by corticosterone.

COMPARISON OF THE CHARACTERISTICS AND OF THE HORMONAL CONTROL OF ENDOMETRIAL AND MYOMETRIAL PROGESTERONE RECEPTORS

1975 ◽  
Vol 66 (3) ◽  
pp. 349-356 ◽  
Author(s):  
M. T. LUU THI ◽  
E. E. BAULIEU ◽  
E. MILGROM
Keyword(s):  
Progesterone Receptor ◽  
Guinea Pig ◽  
Binding Sites ◽  
Guinea Pigs ◽  
Progesterone Receptors ◽  
Relative Increase ◽  
Hormonal Control ◽  
Diploid Genome ◽  
The Absolute ◽  
Hormone Specificity

SUMMARY The characteristics of endometrial and myometrial progesterone receptor of guinea-pig were compared. Affinity for progesterone, hormone specificity, sedimentation properties (in oestrogen-primed animals), inhibition of binding by p-hydroxymercuribenzoate were found to be identical in both tissues. Differences were, however, observed in the hormonal control of the concentration of receptor. In all the situations studied, the concentration of receptors was higher in the endometrium than in the myometrium. In guinea-pigs ovariectomized at dioestrus the concentration was 3500 binding sites per diploid genome in the myometrium and 20300 in the endometrium; 1–3 days after oestradiol injection, this concentration was raised to 46000–38000 and 65000–83000 binding sites respectively. Thus the absolute rise was similar in both tissues but the relative increase was about 15-fold in the myometrium and only three- to fourfold in the endometrium. After injection of 2 mg progesterone, the concentration of receptor previously induced by oestrogen returned to very low values similar to those observed in non-hormonally treated controls. This difference between endometrial and myometrial receptors could be due either to a faster turnover of the latter or to the existence of a stable non-hormonally controlled population of receptors, present only in the endometrium.

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