Gene/Protein Disease Symptom Drug Enzyme Compound
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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)

A comparative radioautographic study of the RNA precursors incorporation on polytene chromosomes of Drosophila in vivo in the cells of salivary glands, and in vitro during incubation of E.coli RNA polymerase on slides with fixed chromosomes was performed.--The pattern of in vivo 3H-uridine incorporation on different sections of the chromosomes drastically differed from the in vitro 3H-UTP incorporation which seems to be much more related to DNA content of the individual small sections. In both cases puffing of the loci resulted in the increase of RNA synthesis but in vitro only 2-3 fold and in vivo much more. Hence, RNA synthesis in vitro was unspecific and did not reflect the in vivo RNA synthesis.--On the other hand, E.coli RNA polymerase completely mimics in vitro the dosage compensation phenomenon making twice as much RNA on one X-chromosome of males (1X2A) as on each of X-chromosomes of diploid (2X2A) and triploid (3X3A) females and super-females (3X2A), and the intermediate amount of RNA on each of X-chromosomes of intersexes (2X3A). It is suggested that the differences in the in vitro template activity of X-chromosomes of cells with different X:A ratio are due to different extent of condensation of their deoxyribonucleoprotein (DNP). Yet, both male and each of female X-chromosomes bind the same amount of thymus histone FI labelled with fluorochrome which indicates that they contain the same amount of "open" regions with exposed chromosomal DNA accessible to external proteins.--On the basis of these observations a hypothesis is put forward which suggests that RNA transcription in animal chromosomes is regulated at two levels by different mechanisms; the first one controls the extent of condensation of DNP of genetic loci and determines their competence to the second mechanism which involves the action of gene-specific activator proteins. According to this hypothesis the phenomenon of dosage compensation of sex-linked genes is due to decondensation of DNP of male X-chromosome which renders its loci twice as responsive to activators as compared to the same loci in females.
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PMID:Comparison of in vivo and in vitro RNA synthesis on polytene chromosomes of Drosophila. 81 77

Methoprene, a chemical analog of juvenile hormone, is toxic when applied to late third-instar larvae of Drosophila melanogaster. Using an ethyl methane sulfonate mutagenesis screen, we have selected two noncomplementing mutants, one of which is nearly 100 times more resistant than wild-type to either methoprene or juvenile hormone III topically applied or incorporated into the diet. The mutation, named methoprene-tolerant (Met), also confers resistance to methoprene-induced pseudotumor formation in larvae as well as to juvenile hormone III- or methoprene-induced vitellogenic oocyte development in adult females. Met adults show little or no cross-resistance to four other insecticides. The mutation was mapped by recombination to a location 35.4 on the X-chromosome and uncovered by chromosomes deficient for the region 10C2-10D4. Complementation was observed between Met and a lethal allele of the RNA polymerase II locus, which is also found in this region. Since the Met mutation also confers resistance to methoprene-induced abnormalities in adult cuticle formation, the autonomy of Met expression could be evaluated in flies mosiac for this mutation. Autonomous expression of Met was found both in abdominal cuticle as well as in external male genitalia. The characteristics of Met are consistent with those expected of a mutant having altered juvenile hormone reception in target tissue.
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PMID:A Drosophila melanogaster mutant resistant to a chemical analog of juvenile hormone. 309 61

The incorporation of 3H-uridine in different regions of polytene chromosomes in live cells of the Drosophila melanogaster salivary glands was compared with the incorporation of 3H-UTP in the same regions under the incubation of cytological preparations of these chromosomes with the E. coli RNA polymerase. The label distribution by regions was compared with the DNA content in them. Individual regions of chromosomes differ by 3H-uridine incorporation in live cells to a much greater extent than by 3H-UTP incorporation in vitro under the incubation with a non-homologous enzyme. RNA synthesis in an exogenous enzyme depends on the DNA content in different chromosome regions to a much greater extent than RNA synthesis in vivo. The correlation of label distribution after 3H-uridine incorporation in live cells and after RNA synthesis in vitro on the preparations by the bacterial RNA polymerase is, correspondingly, very low. This enzyme forms, however, RNA's on puffs 2-3 times more actively than on the same regions in non-puffing state but this difference is dozens of times greater in live cells. RNA synthesis in vitro is, thus, non-specific and does not correspond practically to the intensity of RNA synthesis on the same chromosome regions in live cells. At the same time, as in live cells, the E. coli enzyme synthesizes twice more RNA on the single X-chromosome of males (1X2A) than on each of X-chromosomes of diploid (2X2A) and triploid (3X3A) females or superfemales (3X2A), whereas in intersexes (2X3A) X-chromosomes display intermediate template activity. Thus, RNA synthesis by a heterologous enzyme in vitro does not differ by this index from the synthesis in live cells. It is suggested that differences in the template activity of X-chromosomes in vitro depending on the sex index (X : A) are due to different degree of DNP condensation in these chromosomes. In spite of differences in the degree of condensation, the male X-chromosome binds on the fixed preparation approximately the same amount of thymus histone F1 carrying fluorochrome as each of two female X-chromosomes. Hence, there is no sharp difference between the male and female X-chromosomes by the number and length of DNA regions accessible for interaction with exogenous proteins. On the basis of the data obtained, a hypothesis about two levels and, respectively, two mechanisms of control gene activity in animal chromosomes is considered. The first mechanism is, supposedly, based on decondensation of DNP appears to result in that the same proteins-regulators in the same amount activate corresponding genes in X-chromosome in males twice more strongly than in females.
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PMID:[A comparison of bacterial RNA-polymerase RNA synthesis on polytene Drosophila chromosomes with transcription in living cells]. 421 55

In a search for loci with RNA polymerase related function, we have screened the Drosophila melanogaster genome for regions effective in eliciting a dosage response for total RNA polymerase activity using segmental aneuploids generated by the use of Y-autosome translocations. From this screen we have identified a total of six cytogenetically defined regions which elicit significant dosage response: a single X-chromosome region, 9C-11A, within which resides a known RNA polymerase locus and five noncontiguous regions on chromosome 3 with no previously identified RNA polymerase related function.
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PMID:Detection of cytogenetic regions influencing RNA polymerase activity in Drosophila melanogaster by segmental aneuploid mapping. 644 37

Essential thrombocythemia (ET) is a myeloproliferative disorder characterized by a sustained elevation of the platelet count in the absence of other causes of thrombocytosis. ET is difficult to diagnose, and the demonstration of clonal hematopoiesis may be of value. However, clonality analysis of hematopoietic cells based on the study of the X-chromosome inactivation pattern is complicated by the observation that some normal females present skewed lyonization. Moreover, DNA methylation of X-linked genes in hematopoietic cells may differ from that in other tissues. Appropriate controls for skewed lyonization are therefore critical for the study of clonality. We developed two techniques based on X-chromosome inactivation and polymerase chain reaction (PCR) analysis of polymorphisms, to study clonality in ET patients. Reverse transcriptase-PCR analysis of IDS, P55, and G6PD mRNAs was used to examine the different hematopoietic cell lineages including platelets in patients heterozygous for these polymorphisms and analysis of the HUMARA gene methylation pattern permitted us to study clonality in all nucleated cell fractions of the other patients. Using both types of assay and T lymphocytes as a control tissue for lyonization, clonal hematopoiesis was demonstrated in 28 patients. In 14 patients, the granulocytes were polyclonal; among these patients, platelets were monoclonal in 3 cases, polyclonal in 7 cases, and in the remaining 4 cases this fraction could not be studied because the patients were homozygotes for all RNA markers. No conclusion about clonality could be drawn in 6 cases. Polyclonal hematopoiesis was found in all the cases of reactive thrombocytosis. These findings confirm the high frequency of monoclonal hematopoiesis in ET, the utility of studying platelets, and the possibility of using T lymphocytes as a control tissues for X-chromosome inactivation patterns.
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PMID:Clonality analysis of hematopoiesis in essential thrombocythemia: advantages of studying T lymphocytes and platelets. 897 85

Compartmentalization of the interphase nucleus is an important element in the regulation of gene expression. Here we investigated the functional organization of the interphase nucleus of HeLa cells and primary human fibroblasts. The spatial distribution of proteins involved in transcription (TFIIH and RNA polymerase II) and RNA processing and packaging (hnRNP-U) were analyzed in relation to chromosome territories and large-scale chromatin organization. We present evidence that these proteins are present predominantly in the interchromatin space, inside and between chromosome territories, and are largely excluded by domains of condensed chromatin. We show that they are present throughout the active and inactive X-chromosome territories in primary female fibroblasts, indicating that these proteins can freely diffuse throughout the interchromatin compartment in the interphase nucleus. Furthermore, we established that the in vivo spatial distribution of condensed chromatin in the interphase nucleus does not depend on ongoing transcription. Our data support a conceptually simple model for the functional organization of interphase nuclei.
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PMID:Large-scale chromatin organization and the localization of proteins involved in gene expression in human cells. 1236 63

X-chromosome inactivation (XCI) is highly dynamic during early mouse embryogenesis and strictly depends on the Xist noncoding RNA. The regulation of Xist and its antisense partner Tsix remains however poorly understood. We provide here the first evidence of transcriptional control of Xist expression. We show that RNA polymerase II (RNAPolII) preinitiation complex recruitment and H3 Lys 4 (H3-K4) methylation at the Xist promoter form the basis of the Xist expression profiles that drives both imprinted and random XCI. In embryonic stem (ES) cells, which are derived from the inner cell mass where imprinted XCI is reversed and both Xs are active, we show that Xist is repressed at the level of preinitiation complex (PIC) recruitment. We further demonstrate that Tsix, although highly transcribed in ES cells, is not itself responsible for the transcriptional down-regulation of Xist. Rather, Tsix induces efficient H3-K4 methylation over the entire Xist/Tsix unit. We suggest that chromatin remodeling of the Xist locus induced by biallelic Tsix transcription renders both Xist loci epigenetically equivalent and equally competent for transcription. In this model, Tsix, by resetting the epigenetic state of the Xist/Tsix locus, mediates the transition from imprinted to random XCI.
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PMID:Tsix transcription across the Xist gene alters chromatin conformation without affecting Xist transcription: implications for X-chromosome inactivation. 1596 97

CTCF is a ubiquitous transcription factor that is involved in numerous, seemingly unrelated functions. These functions include, but are not limited to, positive or negative regulation of transcription, enhancer-blocking activities at developmentally regulated gene clusters and at imprinted loci, and X-chromosome inactivation. Here, we review recent data acquired with state-of-the-art technologies that illuminate possible mechanisms behind the diversity of CTCF functions. CTCF interacts with numerous protein partners, including cohesin, nucleophosmin, PARP1, Yy1 and RNA polymerase II. We propose that CTCF interacts with one or two different partners according to the biological context, applying the Roman principle of governance, 'divide and rule' (divide et impera).
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PMID:CTCF and its protein partners: divide and rule? 1938 94

Non-coding (nc)RNAs are key players in numerous biological processes such as gene regulation, chromatin domain formation and genome stability. Large ncRNAs interact with histone modifiers and are involved in cancer development, X-chromosome inactivation and autosomal gene imprinting. However, despite recent evidence showing that pervasive transcription is more widespread than previously thought, only a few examples mediating gene regulation in eukaryotes have been described. In Saccharomyces cerevisiae, the bona-fide regulatory ncRNAs are destabilized by the Xrn1 5'-3' RNA exonuclease (also known as Kem1), but the genome-wide characterization of the entire regulatory ncRNA family remains elusive. Here, using strand-specific RNA sequencing (RNA-seq), we identify a novel class of 1,658 Xrn1-sensitive unstable transcripts (XUTs) in which 66% are antisense to open reading frames. These transcripts are polyadenylated and RNA polymerase II (RNAPII)-dependent. The majority of XUTs strongly accumulate in lithium-containing media, indicating that they might have a role in adaptive responses to changes in growth conditions. Notably, RNAPII chromatin immunoprecipitation followed by DNA sequencing (ChIP-seq) analysis of Xrn1-deficient strains revealed a significant decrease of RNAPII occupancy over 273 genes with antisense XUTs. These genes show an unusual bias for H3K4me3 marks and require the Set1 histone H3 lysine 4 methyl-transferase for silencing. Furthermore, abolishing H3K4me3 triggers the silencing of other genes with antisense XUTs, supporting a model in which H3K4me3 antagonizes antisense ncRNA repressive activity. Our results demonstrate that antisense ncRNA-mediated regulation is a general regulatory pathway for gene expression in S. cerevisiae.
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PMID:XUTs are a class of Xrn1-sensitive antisense regulatory non-coding RNA in yeast. 2169 27

The X-chromosome gene regulatory process called dosage compensation ensures that males (1X) and females (2X) express equal levels of X-chromosome transcripts. The mechanism in Caenorhabditis elegans has been elusive due to improperly annotated transcription start sites (TSSs). Here we define TSSs and the distribution of transcriptionally engaged RNA polymerase II (Pol II) genome-wide in wild-type and dosage-compensation-defective animals to dissect this regulatory mechanism. Our TSS-mapping strategy integrates GRO-seq, which tracks nascent transcription, with a new derivative of this method, called GRO-cap, which recovers nascent RNAs with 5' caps prior to their removal by co-transcriptional processing. Our analyses reveal that promoter-proximal pausing is rare, unlike in other metazoans, and promoters are unexpectedly far upstream from the 5' ends of mature mRNAs. We find that C. elegans equalizes X-chromosome expression between the sexes, to a level equivalent to autosomes, by reducing Pol II recruitment to promoters of hermaphrodite X-linked genes using a chromosome-restructuring condensin complex. DOI:http://dx.doi.org/10.7554/eLife.00808.001.
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PMID:Condensin controls recruitment of RNA polymerase II to achieve nematode X-chromosome dosage compensation. 2379 97


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