DIFFERENTIAL DISTRIBUTION OF OESTROGEN RECEPTORS IN SUBFRACTIONS OF OVIDUCT CHROMATIN

1977 ◽  
Vol 84 (1) ◽  
pp. 215-224 ◽  
Author(s):  
Kari Hemminki

ABSTRACT DNase II shearing and MgCl2 precipitation of chick oviduct chromatin were applied to prepare fractions which resembled template-active and template-inactive chromatin fraction as described for other tissues. The labelling of these preparations with [3H]oestradiol was studied in short-term tissue cultures. The MgCl2 soluble (∼ template-active) chromatin bound 1.8 times more oestradiol than the MgCl2 insoluble chromatin when immature chicks were used. The proportion was 4.3-fold in oestrogen stimulated chicks and 4.6-fold in laying hens. Similar results were obtained with an oestrogen exchange assay which measured the concentration of endogenous receptors. These data suggest that hormone stimulation is accompanied by a selective activation of oviduct chromatin leading to an enrichment of acceptor sites for oestradiol receptors in the template-active chromatin fraction.

1978 ◽  
Vol 173 (1) ◽  
pp. 155-164 ◽  
Author(s):  
R W Jones

The histone compositions of a chromatin fraction containing ribosomal DNA and of the remaining macronuclear chromatin of Tetrahymena pyriformis was analysed by gel electrophoresis. These chromatin fractions were used as models for transcriptionally active and inactive chromatin respectively. The extent of histone modification, as indicated by the distribution of histone between differently charged subspecies in acid-urea gels, is not grossly different in the two chromatin fractions. However, histone H1 is present, but may be differently modified in the two chromatin fractions. The histone/DNA ratio in ribosomal chromatin, measured after sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of samples of chromatin, was found to be the same whether chromatin was extracted from growing or stationary organisms, and to be approx. 40% of this ratio in the remaining macronuclear chromatin. The implications of these results for the possible structure of transcriptionally active chromatin are discussed.


1997 ◽  
Vol 322 (1) ◽  
pp. 273-279 ◽  
Author(s):  
Kulbhushan TIKOO ◽  
Sunita GUPTA ◽  
Q. Anwar HAMID ◽  
Vanya SHAH ◽  
Bishwanath CHATTERJEE ◽  
...  

Rat liver nuclei were isolated in low-ionic-strength buffer in the absence of bi- and multi-valent cations. Digestion of these nuclei by endogenous nuclease, micrococcal nuclease and DNase I revealed that a minor chromatin fraction was preferentially digested into poly- and oligo-nucleosomes. Southern blot hybridization with various active gene probes confirmed that these chromatin fragments represent coding and 5ƀ upstream regions of transcriptionally active chromatin. Active chromatin fragments were released selectively into the medium, with inactive chromatin remaining inside the nuclei, under the above ionic conditions. The inclusion of bivalent cations during the digestion of nuclei reversed the solubility behaviour of active chromatin. Rearrangement and exchange of histone H1 between chromatin fragments was prevented by using low-salt conditions in all steps in the absence of bivalent cations. All histones, including H1, were present in stoichiometric amounts in this active chromatin fraction. Active nucleosomes showed a lower electrophoretic mobility than bulk nucleosomes in an acrylamide/agarose composite gel in the absence of Mg2+, but were selectively bound to the gel in the presence of this ion.


1986 ◽  
Vol 113 (3) ◽  
pp. 346-354
Author(s):  
Annalisa Tanini ◽  
Maria Luisa Brandi ◽  
Umberto Modigliani ◽  
Carlo M. Rotella ◽  
Roberto Toccafondi

Abstract. TSH-induced cAMP accumulation in cells obtained from normal and pathological thyroid tissue was studied during the first 12 days of primary culture. In normal thyroid tissue cultures (N = 7), the response of cAMP to TSH was present from the second day of culture and reached its maximum after 8 days. A similar behaviour was observed in cultures obtained from euthyroid sporadic goitres (N = 8), even if the rate of response was slightly lower than that of normal tissue. Similarly, cultured cells from euthyroid 'autonomous' nodules (N = 8) appeared to be responsive to TSH during the period of study, but the rate of response was also lower than in the controls. On the contrary, in cultures obtained from toxic adenomas (N = 5) and from diffuse toxic goitres (N = 5) the response to TSH was absent during the first 4 days of culture. The cells became sensitive to TSH from 6 and 6 day onwards, with the rate of response increasing progressively and reaching its maximum on day 12. Finally, in cultured cells obtained from different areas of multinodular toxic goitres (N = 4), the response to TSH was similar to that of euthyroid goitres in cells prepared from 'cold' areas, and to that of toxic adenomas in cells obtained from 'hot' areas. The present data demonstrate the existence of an inhibitory action of unknown factors, possibly iodothyronines or thyroglobulin, on the TSH effect in short-term cultures obtained from thyrotoxic tissues. A normal TSH responsiveness can be restored when the culture is prolonged.


1999 ◽  
Vol 112 (20) ◽  
pp. 3537-3548 ◽  
Author(s):  
C. Pittoggi ◽  
L. Renzi ◽  
G. Zaccagnini ◽  
D. Cimini ◽  
F. Degrassi ◽  
...  

We have characterized a nuclease hypersensitive chromatin fraction from murine spermatozoa. Endogenous nuclease activity can be induced in mouse epididymal spermatozoa by appropriate stimuli and cause the localized degradation of chromosomal DNA. Based on these observations, we have isolated nuclease hypersensitive chromatin regions released from spermatozoa in the supernatant of pelleted sperm cells, and have cloned and characterized the DNA. Gel electrophoresis of end-labelled released DNA fragments showed a typical nucleosomal distribution. Peripherally distributed nucleohistones were visualized by immunofluorescence in sperm nuclei, and histones were identified by western blot in sperm chromatin. Moreover, the released DNA is enriched in retroposon DNA from a variety of families. FISH and immunofluorescence analysis showed that retroposon DNA and nucleohistone chromatin co-localize and are both peripherically distributed in nuclei of spermatozoa. In contrast, a major satellite DNA probe, used for control, co-localizes with highly condensed chromatin in the central region of sperm nuclei. The nuclear Ran and RCC1 proteins were also visualized in the dorsal margin of sperm nuclei, and were abundantly released with the hypersensitive chromatin fraction. Together, these results indicate that nucleohistone chromatin fraction(s) with typical features of ‘active’ chromatin are present in murine spermatozoa, are hypersensitive to nuclease cleavage, enriched in retroposon DNA and organized in nucleosomal domains. These observations suggest that nucleohistone domains identify a fraction of the sperm genome which may be functional during early embryogenesis.


Author(s):  
Claudio Festuccia ◽  
Mauro Bologna ◽  
Carlo Vicentini ◽  
Antonella Tacconelli ◽  
Roberto Miano ◽  
...  

1990 ◽  
Vol 271 (1) ◽  
pp. 67-73 ◽  
Author(s):  
M J Hendzel ◽  
J R Davie

The incorporation of newly synthesized histones among various chromatin fraction isolated from non-replicating cell-cycle-phase-Go chicken immature erythrocytes was investigated. We find that newly synthesized erythroid-specific histone Hl variant H5, is incorporated randomly into chromatin. In contrast, newly synthesized nucleosomal histones H2A, H2A.Z, H2B, H3.3, and H4 are preferentially found in a fraction that is highly enriched in active/competent gene chromatin fragments and depleted in repressed gene chromatin. Moreover, ubiquitinated species of histones H2A and H2B and hyperacetylated species of H4 and H2B, which are complexed to active DNA, are labelled. These observations provide evidence that newly synthesized histones preferentially exchange with the nucleosomal histones of transcriptionally active/component chromatin domains. The results of this study suggest that nucleosomes of active chromatin may be inherently less stable than bulk nucleosomes in vivo and have implications for chromatin remodelling.


1993 ◽  
Vol 55 (4) ◽  
pp. 672-677 ◽  
Author(s):  
C. Van Pachterbeke ◽  
M. Tuynder ◽  
J. Rommelaere ◽  
J. P. Cosyn ◽  
L. Lespagnard ◽  
...  

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