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Query: EC:2.7.7.6 (RNA polymerase)
34,946 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The abilities of H1 and H5 histones from immature and mature pigeon erythroid cells and subfractions of H5 histones with different content of alkali-labile phosphorus to restrict RNA synthesis were compared in an in vitro transcription system with bacterial RNA polymerase. It has been found that: a) H1 histones from both sources exhibit similar efficiency. b) H5 histones from immature cells restrict transcription to a lesser extent than the same histone from the mature erythrocyte. c) The subfraction of H5 histone from immature cells with the higher content of alkali-labile phosphorus restrict transcription less than the subfraction with the lower degree of phosphorylation. The results suggest that H5 histone, which first appears in the erythroblasts in a highly phosphorylated form, does not display its potential 'repressor' properties. During erythrocyte maturation H5 histone is dephosphorylated and this causes the massive inactivation of erythrocyte genome.
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PMID:Existence of differences in repressor properties between serine-rich histones (H5, F2c) from immature and mature pigeon erythroid cells. 64 Mar 4

Hg-UMP-containing transcripts made from chick erythroid chromatins with E. coli RNA polymerase hybridize to chick globin cDNA. Contamination with endogenous globin RNA has been largely removed by purification on SH-agarose columns at 55 degrees C. Some endogenous globin mRNA sequences remain, probably as hybrids with "anti-sense" Hg-transcripts produced by RNA-dependent RNA synthesis. Heating to 115 degrees C before SH-agarose chromatography eliminates these contaminants. Hg-transcripts from adult and embryonic erythroid chromatins purified by this method are hybridized to globin cDNA; they contain a 4- to 6-fold higher proportion of globin-specific sequences (10-13 PPM) than do transcripts from brain chromatin. Dissociation of erythroid chromatins in salt and urea, followed by reconstitution using standard methods, destroys even this low degree of specificity.
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PMID:Is there specific transcription from isolated chromatin? 65 20

In order to provide more information on nucleolar changes occurring during cell differentiation and maturation, pigeon erythroid cells have been studied by means of a simple light microscopic cytochemical procedure for the demonstration of the RNA containing structures and with conventional transmission electron microscopy. Nucleoli with more or less distinct nucleolonemata present in early erythroblasts were replaced by ring-shaped nucleoli and, finally, by micronucleoli in more mature erythroid cells. In contrast to the previously studied chick embryos, chickens and hens which possess micronucleoli in almost all mature erythrocytes, mature pigeon erythrocytes are mostly without any nucleoli. The ultrastructural organization of nucleoli with more or less distinct nucleolonemata and ring-shaped nucleoli do not show differences, in comparison with such forms of nucleoli in other cells. Micronucleoli in pigeon erythroblasts are characterized by degranulation, suggesting the inhibition of the formation of the nucleolar granular components. Some nucleoli show a segregation of the nucleolar components, indicating the inactivation of the nucleolar RNA polymerase, and occasionally chromatin retraction from the nucleolar body, suggesting the loss of the template. In mature erythrocytes, micronucleoli consist mainly of fibrillar components, and the perinucleolar chromatin is partially retracted from the nucleolar body of such nucleoli.
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PMID:Studies on nucleoli of pigeon erythroid cells. 68 48

Chromosomal non-histone proteins are obtained from nuclei of two types of pigeon erythroid cells: erythroblasts (cells active in RNA synthesis) and erythrocytes (cells with repressed RNA synthesis). They are well soluble in solutions of low ionic strength. Electrophoretic separation of the obtained non-histone proteins in polyacrylamide gels with urea and SDS shows the presence of qualitative differences in the pattern of non-histone proteins of chromatine from erythroblasts and erythrocytes. By electrophoresis in urea some protein bands of non-histone proteins of chromatine from erythroblasts were found which disappear with the aging of cells. At the same time two protein fractions were observed in chromatine from erythrocytes which were absent in that of erythroblasts. Disappearance of some high molecular weight protein fractions from erythrocyte chromatine as compared to erythroblasts was observed by separation of the non-histone proteins in the presence of SDS. These fractions of the non-histone proteins disappearing during aging of cells are well extractable from erythroblast chromatine by 0.35 M NaCl solution. In the in vitro system with E. coli RNA polymerase addition of non-histone proteins of chromatine from erythroblasts to chromatine from erythrocytes increases RNA synthesis 2--3 times. At the same time addition of non-histone proteins from erythrocytes is either without any influence on this process or somewhat inhibiting.
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PMID:[Comparative investigation of the non-histone proteins of chromatin from pigeon erythroblasts and erythrocytes]. 80 71

Avian erythroid cells were separated into five developmental stages by sedimentation on discontinuous isotonic albumin gradients. Solubilized enzyme activities from whole cells were partially purified and characterized by ion exchange and ion filtration chromatography and velocity sedimenttation analysis. Three nucleotide polymerase types were investigated: (a) DNA-dependent RNA polymerases; (b) RNA-dependent terminal ribonucleotidyltransferases, and (c) DNA-dependent DNA polymerases. The two characteristic forms of eucaryotic DNA-dependent RNA polymerases, polymerase I (nucleolar) and polymerase II (nucleoplasmic), were identified. Polymerase III was only marginally detectable even in the earliest developmental populations. At least two species of RNA-dependent terminal ribosyltransferases were present. One apparently was the poly(A) polymerase observed in other systems. The other terminal transferase was present in two chromatographic forms, required an RNA primer, and used UTP and/or CTP as particularly efficient substrates. Three DNA polymerase activities were resolved, two of which were characteristic of the alpha and beta DNA polymerases described in other eucaryotic systems. The third polymerase was not the gamma polymerase but a separate entity. Poly(dC)-dependent RNA polymerase activity, associated with the alpha polymerase, was relatively enriched in the third DNA polymerase species. The activity levels of the nucleotide polymerases were monitored as a function of red cell maturation. Characteristic declining patterns of activity were obtained for each enzyme which correlate well with the synthetic rates of their in vivo products where these are known. These results correlate well with the synthetic rates of their in vivo products where these are known. These results are consistent with the postulate that the general transcriptive and replicative control processes operating during development may involve changes in the level of the requisite polymerases.
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PMID:Nucleotide polymerases in the developing avian erythrocyte. 83 21

Globin gene switching in sheep and goats has been used as a model system for examining gene expression in differentiating red blood cells. Sheep and goats switch from the synthesis of hemoglobin A to hemoglobin C in response to erythropoietin. The regulatory mechanism producing this switch in hemoglobin types could occur at the cellular, nuclear, or cytoplasmic level. Evidence is presented which suggests that regulation is occurring, in fact, at the nuclear level. Sheep and goat erythroid colonies have been grown in plasma clot culture in order to study the synthesis of individual globin chains. Erythropoietin is required for colony formation. The switch from hemoglobin A to hemoglobin C synthesis requires not only colony formation but also a higher concentration of erythropoietin than is required just for the production of colonies. A cell-free transcriptional system using bone marrow chromatin and mammalian DNA-dependent RNA polymerase has been developed in order to examine the nuclear control mechanisms in more detail.
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PMID:Activation and inactivation of genes determining hemoglobin types.20s. 116 4

We have used the globin family of genes in chicken to study developmental regulation of gene expression, both at the level of individual interaction of trans-acting factors with local promoters and enhancers, and at the level of chromatin structure. Regulation of all members of the alpha- and beta-globin clusters is affected by the erythroid regulatory factor GATA-1. Separate mechanisms exist for regulation of individual members of the family. As an example, we describe the control mechanisms that play a role in the expression of the rho-globin gene, which is expressed only in primitive lineage erythroid cells. In addressing the involvement of chromatin structure in gene activation, we have examined the role of locus control elements, and also considered the way in which RNA polymerase molecules might accommodate to the presence of nucleosomes on transcribed genes.
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PMID:Developmental regulation of globin gene expression. 129 48

In this study we describe DNA-RNA complexes in matrix DNA of Friend cells. The presence of such unusual structures is confirmed by the following evidence. When a preparation of matrix DNA is electrophoresed in agarose an RNA component always migrates together with DNA. There should be a close interaction between DNA and RNA in such a preparation because the presence of the RNA component causes resistance of DNA to DNase I and Exo III. An intimate, hybrid-type association of part of the RNA component with DNA is indicated also by the fact that about 20% of this RNA is sensitive to RNase H. By specific inhibition of the RNA synthesis with alpha-amanitin and actinomycin D it was shown that the bulk of associated RNA is transcribed by RNA polymerase III. Hybridization experiments showed similarity between the DNA sequences isolated from the complexes and those from the base of dehistonized DNA loops obtained by high-salt extraction of nuclei. This observation suggests that the complexes might represent attachment sites of nuclear DNA to the matrix: possibly, the attachment is mediated via the RNA component. Experiments with induction of erythroid differentiation indicated that a profound reorganization of the nucleus, accompanying terminal differentiation, leads to a striking reduction in the number of complexes and thus in the number of attachment sites. This suggests that the complexes should function as transient attachment sites.
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PMID:DNA-RNA complexes that might represent transient attachment sites of nuclear DNA to the matrix. 169 80

Feline leukemia virus, subgroup C/Sarma (FeLV-C/Sarma) induces pure red blood cell aplasia in cats. Although erythroid (BFU-E and CFU-E) and granulocyte/macrophage (CFU-GM) progenitors are infected with this virus, only erythropoiesis is impaired. Two to 3 weeks before the onset of anemia, CFU-E become undetectable in marrow cultures while earlier erythroid progenitors (BFU-E) persist, suggesting that FeLV-C/Sarma (presumably via its envelope glycoprotein gp70) inhibits the differentiation of BFU-E to CFU-E in vivo. To correlate in vitro observations with the progression of disease, prospective studies were performed in six cats. These studies showed that at the time that the frequencies of CFU-E decreased in marrow cultures, BFU-E no longer responded to hematopoietic growth factor(s), although the responses of CFU-GM were unchanged. In further studies, anemic cats received suramin, a reverse-transcriptase inhibitor with other diverse effects. Within 4 to 14 days, erythropoiesis improved and up to 1,616 CFU-E were detected per 10(5) marrow mononuclear cells. However, progenitor cells remained infected, suggesting that suramin modulated erythroid differentiation without inhibiting progenitor infection. These observations led to the hypothesis that the gp70 of FeLV-C/Sarma impairs BFU-E differentiation by interference with ligand/receptor interactions or signal transduction pathways unique to erythroid cells. Understanding this mechanism should provide insights into the interactions controlling early erythropoiesis.
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PMID:Retrovirus-induced feline pure red blood cell aplasia: pathogenesis and response to suramin. 184 31

The amounts of mouse beta major-globin mRNA and globin gene upstream RNA polymerase III transcripts were compared during the differentiation of purified erythroid colony-forming progenitor cells (CFU-E) in vitro. The accumulation of each RNA was determined relative to the 100% levels in fetal liver erythroid cells. The mRNA level was low in freshly isolated CFU-E and began to accumulate only after a lag period of approximately 2-4 h. It continued to accumulate for approximately 40 h thereafter, at which point it was comparable to that in the fetal liver. In contrast, in three of four CFU-E preparations, the relative level of upstream RNAs was high in freshly isolated CFU-E and reached the maximal level (defined as 100% of the fetal liver level) more rapidly than did the mRNA. The early induction and accelerated accumulation of upstream RNAs in immature erythroid cells suggest some role for these RNAs at an early stage of globin gene activation.
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PMID:Expression of the 5'-flanking region of the beta major-globin gene in cultured murine erythroid precursor cells. 244 25


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