Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.31.1 (micrococcal nuclease)
 2,818 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

As an approach to the mechanism of the nuclear translocation of estrogen receptor, the estradiol nuclear receptor (RN) of lamb endometrium was extracted with micrococcal nuclease at 2--4 degrees and compared to the "native" 8S and to the Ca2+-transformed cytosol receptors. After extensive digestion of chromatin, giving up to 10% perchloric acid-soluble DNA and a majority of nucleosome monomers, up to 80% of the RN was extracted and under low ionic strength. This RN was found to be completely different from the partially proteolyzed Ca2+-transformed cytosol receptor. It migrated with a sedimentation constant of 4 and 6 S. The Stokes radius of the predominant form as determined by ACA 34 chromatography was 5.3 nm. The calculated apparent molecular weights were 130,000 and 90,000, respectively. The RN was able to bind DNA and was eluted from a diethylaminoethyl cellulose column at 0.23 and 0.30 M KCl. We conclude that the mechanism proposed by Puca et al., according to which the Ca2+-transformed cytosol receptor is split by a Ca2+ receptor-transforming factor into a smaller form able to cross the nuclear membrane, is very unlikely.
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PMID:Comparison between different forms of estrogen cytosol receptor and the nuclear receptor extracted by micrococcal nuclease. 69 61

L-Triiodothyronine induces malic enzyme in explants from human adipose tissue. Consequently, we looked for the presence of receptors for L-triiodothyronine in nuclei isolated from human adipose tissue. The binding of 125I-triiodothyronine by the nuclei was time- and temperature-dependent. At 37 degrees C binding reached a steady state after 60 minutes. Dithiothreitol enhanced total binding and suppressed nonspecific binding. Scatchard analysis showed the presence of a single class of binding sites. The apparent association constant, Ka, was 0.13 +/- 0.03 X 10(10) M-1, the maximal binding capacity 2.20 +/- 0.81 pmol/mg DNA (mean +/- SD, n = 7) and the number of binding sites 8,000/nucleus. L-Triiodothyronine and D-triiodothyronine had equal affinity to the nuclear receptor; triiodothyroacetic acid had three times higher affinity. L- and D-thyroxine had 8% and 12%, respectively, and tetraiodothyroacetic acid had 19% affinity compared to that of L-triiodothyronine. Reverse triiodothyronine was a weak competitor. Digestion of nuclei with micrococcal nuclease abolished specific binding. These results show that nuclei from human white adipose tissue possess high affinity receptors for L-triiodothyronine, which are associated with nuclear chromatin. It is likely that induction of malic enzyme in human adipose tissue by L-triiodothyronine is mediated by the nuclear receptors.
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PMID:Nuclear triiodothyronine receptor of human adipose tissue. 334 8

High affinity-low capacity binding sites for thyroid hormone have been identified in the nuclei of glial (C6) and neuronal (Neuro 2A) cultured cells. Equilibrium dissociation constants, determined by Scatchard analysis, were very similar in both types of cells (0.2-0.3 nM). The relative affinity of hormonal analogs was also similar: the affinity for T3 was lower than for triiodothyroacetic acid and higher than for T4 or tetraiodothyroacetic acid. The sedimentation coefficients obtained by gradient centrifugation of nuclear receptor extracted with 0.4 M KCl or excised by micrococcal nuclease digestion were 3.5 S and 6.5 S, respectively. These results suggest that the thyroid hormone receptor is not restricted to neuronal cells, but also appears in cells of glial origin.
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PMID:Identification and characterization of L-triiodothyronine receptors in cells of glial and neuronal origin. 376 67

The thyroid hormone receptor is a chromatin-associated protein which appears to mediate the actions of the thyroid hormones in mammalian cells. Unlike steroid hormone receptors, a cytoplasmic form of the receptor has not been identified, and the factors which govern the nuclear concentrations of the receptor are poorly understood. Using cultured GH1 cells, a rat pituitary cell line, we having previously demonstrated that thyroid hormones reduces the concentration of its receptor by a mechanism which involves the association of the ligand with the receptor binding site (Samuels, H.H., Stanley, F., and Shapiro, L.E. (1977) J. Biol. Chem. 252, 6052-6060). In this study, we demonstrate that n-butyrate and other aliphatic carboxylic acids elicit a reduction of thyroid hormone nuclear receptor levels without altering total cell protein synthetic rates. In contrast, the nuclear association and total cell level of the glucocorticoid receptor is not altered by n-butyrate. Evidence is presented that the aliphatic carboxylic acid-mediated reduction of thyroid hormone nuclear receptor levels is secondary to the inhibitory effect of these compounds on chromatin-associated deacetylases which is reflected as an increase in the acetylation of the nucleosome core histones. Isokinetic gradient centrifugation of chromatin solubilized from GH1 cell nuclei by micrococcal nuclease indicates that the receptor exists as a form associated with high molecular weight chromatin, as a 12.5 S form that sediments slightly faster than the bulk of the mononucleosomes, and as a 6.5 S form which appears to remain associated with low molecular weight chromatin components. Exclusive of the receptor associated with the high molecular weight chromatin, the 6.5 S form represents 80% and the 12.5 S form 10% of the receptor resolved in the gradient. n-Butyrate decreases both forms to the same degree suggesting that they are generated from the same "entity" of chromatin structure. Studies on the reappearance of receptor after restoration of the chromatin to the "normal" acetylated state are consistent with a model in which the affinity of chromatin for newly synthesized receptor is diminished in the "hyperacetylated" state.
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PMID:Thyroid hormone nuclear receptor levels are influenced by the acetylation of chromatin-associated proteins. 624 82

We have investigated the association of the triiodothyronine (T3) nuclear receptor with rat liver chromatin by the use of selective endonuclease digestion and differential solubilization. The T3 receptor was found in a fraction of chromatin having some of the characteristics of active chromatin: (a) It is highly sensitive to DNase I and micrococcal nuclease digestion; (b) it is enriched in nonhistone proteins and depleted of histone (H-1). Micrococcal nuclease and pancreatic DNase I excised two receptor-containing fragments from chromatin, a minor (12--14 S) form and a major (5.5--6.0 S) form. The latter structure has a Stokes radius of 42 +/- 2 A and an estimated molecular weight of 95400 when a partial specific volume of 0.725 cm3/g for protein is used. It contains DNA but no histones. Similar receptor-containing fragments were excised from chromatin of other rat tissues, including brain, kidney, and heart. Both the 5.5--6 S and the 12--14 S receptor-containing chromatin structures are converted by 0.5 M KCl to the 3.5 S form (R0 35 A molecular weight 50500). Titration with micrococcal nuclease and pancreatic DNase I revealed that the 5.5--6 S form is preferentially excised by endonuclease. Neither receptor occupancy nor thyroidal status had an apparent effect on the susceptibility of chromatin to endonucleolytic digestion nor did they influence the distribution of T3 receptors in chromatin. Our results suggest that T3 receptors are not tightly associated with nucleosomes, the basic subunit of chromatin, but are associated with the DNA-linking nucleosomes in structurally modified regions of chromatin in rat liver nuclei. The T3 receptor-containing fragment may well represent a higher order of structural complexity necessary for T3 action at the cellular level.
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PMID:Association of the thyroid hormone receptor with rat liver chromatin. 627 79

Transcription of the retinoic acid receptor beta2 (RARbeta2) gene is induced by retinoic acid (RA) in mouse P19 embryonal carcinoma (EC) cells. Here we studied RA-induced chromatin structure alterations in the endogenous RARbeta2 promoter and in an integrated, multicopy RARbeta2 promoter in EC cells. RA markedly increased restriction site accessibility within the promoter, including a site near the RA responsive element (RARE) to which the nuclear receptor retinoid X receptor (RXR)-RAR heterodimer binds. These changes coincided with RA-induced alterations in the DNase I hypersensitivity pattern in and around the promoter. These changes became undetectable upon removal of RA, which coincided with the extinction of transcription. Analyses with receptor-selective ligands and an antagonist showed that increase in restriction site accessibility correlates with transcriptional activation, which parallels the RA-induced in vivo footprint of the promoter. Despite these changes, the micrococcal nuclease digestion profile of this promoter was not altered by RA. These results indicate that concurrent with the binding of the RXR-RAR heterodimer to the RARE, the local chromatin structure undergoes dynamic, reversible changes in and around the promoter without globally affecting the nucleosomal organization.
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PMID:Retinoid-induced chromatin structure alterations in the retinoic acid receptor beta2 promoter. 934 11