Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ovaries of the amphibian Triturus viridescens contain a considerable amount of 7 to 12 S RNA which fractionates with poly(A)+ RNA on oligo(dT)-cellulose column chromatography and directs the synthesis of all five classes of histones in a wheat germ cell-free system. The polyadenylate tracts associated with this 7 to 12 S poly (A)+ RNA are heterogeneous in length, ranging from approximately 60 to 120 nucleotides. Partially purified subfractions of histone mRNA templates were isolated from this RNA by preparative polyacrylamide gel electrophoresis. The correlation between the poly(A) content, the template activity in vitro, and the cell-free products encoded by these RNA subfractions suggests that Triturus ovary RNA contains poly(A)+ histone mRNA. The 7 to 12 S poly(A)+ RNA, but not 7 to 12 S poly(A)- RNA, is an effective template for the avian myeloblastosis virus reverse transcriptase-directed synthesis of complementary DNA (cDNA) in the presence of oligo(dT) primer. In the absence of primer, virtually no cDNA is synthesized. When cDNA complementary to 7 to 12 S poly(A)+ RNA is hybridized in situ to Drosophila melanogaster polytene chromosome preparations. cDNA hybrids are found primarily in the region 39D-E, the locus of the histone genes in Drosophila. This cDNA also hybridizes to the sea urchin histone gene sequences contained in the chimeric bacterial plasmids pSp2 and pSp17. These results provide strong evidence for the reality of polyadenylated histone mRNA in Triturus ovary.
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PMID:A portion of all major classes of histone messenger RNA in amphibian oocytes is polyadenylated. 56 2

The organization of the genes coding for histones in the chicken has been examined, with special reference to that coding for the tissue-specific, developmentally regulated histone H5. Two recombinant phages containing sequences complementary to cloned H5 cDNA have been isolated from a genomic chicken library. The clones have been characterized by heteroduplex formation, restriction nuclease analysis, hybridization to cloned homologous histone gene probes, and DNA sequencing. Hybridization to genomic DNA has shown that there is only one copy of the H5 gene per haploid genome, whereas there are six to 11 copies of the genes for the other histones. Examination of 29 X 10(3) base-pairs of DNA sequences flanking the H5 gene has revealed the absence of any other histone genes which, although not tandemly reiterated, for the most part appear to reside in loosely organized clusters. The complete DNA sequence of the H5 gene and flanking regions, as well as the mapping of the 5'-end of its messenger RNA by primer extension with AMV reverse transcriptase, has shown that the gene has no introns and little homology to other histone genes, including those for H1.
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PMID:Genomic organization of the genes coding for the six main histones of the chicken: complete sequence of the H5 gene. 631 52

Histone mRNA from S. cerevisiae has been identified and partially purified. The RNA is quantitatively retained on oligo (dT) cellulose or poly(U) sepharose as assayed by in vitro translation or hybridization of radiolabelled cloned yeast histone sequences to RNA immobilized on DBM paper. Retention of yeast histone mRNA on either of these chromatographic systems is most likely the result of polyadenylation since, when primed with oligo (dT), the RNA is an extremely good template for reverse transcriptase, as determined by hybrid arrest translation or by hybridization to D. melanogaster histone DNA sequences.
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PMID:Yeast histone mRNA is polyadenylated. 700 28

To determine the effects of acute myocardial infarction on the expression of insulin-like growth factor1 (IGF1) and insulin-like growth factor1 receptors (IGF-1R) on the surviving myocytes of the left and right ventricles, large infarcts were produced in rats and the animals sacrificed 2 days later. Hemodynamic measurements of left and right ventricular pressures, +dP/dt and -dP/dt, and central venous pressure documented that coronary occlusion was associated with a severe impairment of cardiac function. By employing reverse transcriptase polymerase chain reaction (RTPCR), a low level of expression of IGF-1R mRNA was detected in myocytes from sham-operated rats. Acute myocardial infarction was found to enhance by nearly twofold the message for IGF-1R in viable myocytes biventricularly. Moreover, IGF1 mRNA increased 4.3-fold and 9.4-fold in left and right myocytes, respectively. In order to establish whether the upregulation of IGF1 and IGF-1R with infarction was coupled with induction of late growth related genes, which are known to be implicated in DNA replication and mitotic division, proliferating cell nuclear antigen (PCNA) and histone-H3 expression was assessed by Northern blot and RTPCR. The level of expression of PCNA mRNA was found to be increased 3.9-fold and 2.4-fold in left and right myocytes, respectively from infarcted hearts. Corresponding increments in histone-H3 mRNA were 25.5-fold and 5.3-fold, respectively. However, PCNA protein as detected by immunoperoxidase staining was restricted to a limited number of myocyte nuclei adjacent to the necrotic myocardium of the left ventricle. In conclusion, acute myocardial infarction is associated with enhanced expression of IGF1 and IGF-1R on stressed myocytes, and this phenomenon may activate genes essential for DNA synthesis, possibly affecting myocyte growth. These processes may be fundamental for the reconstitution of tissue mass and amelioration of function after infarction.
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PMID:Upregulation of IGF1, IGF1-receptor, and late growth related genes in ventricular myocytes acutely after infarction in rats. 750 76

Repetitive sequences with oligo A tails were observed in Dra1 fragments of Bombyx mori genomic DNA. The full sequence of the element, an abundant non-LTR retrotransposon of B. mori, was determined by assembling inner restriction fragments. This element, designated L1Bm, contained two ORFs encoding a gag-like protein and reverse transcriptase (RT), respectively. An endonuclease domain was identified at the N-terminus of the RT sequence. The homology search of the amino acid sequences revealed that L1Bm belongs evolutionally to the same family as various other non-LTR retrotransposons of Drosophila and Mosquito. On the other hand, L1Bm resembles the L1 element of human genome in its high copy number and its frequent truncation at the 5'-side. Some units of the histone gene repeat in B. mori possess complete L1Bm elements in a 3'-flanking region of H2b.
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PMID:A major non-LTR retrotransposon of Bombyx mori, L1Bm. 930 19

The stimulatory effects of several DNA-binding basic proteins (histone and protamine) and HIV-1 Rev with arginine (Arg)-rich clusters on the activity of casein kinase II (CK-II) were investigated in vitro. It was found that recombinant Rev (rRev) and the synthetic oligo-fragments corresponding to the amino acid sequences of its Arg-rich cluster stimulate CK-II activity in a dose-dependent manner. The activated CK-II phosphorylates several cellular and viral proteins in HIV-1 infected human MOLT-4 cells, and also phosphorylates HIV-1 structural proteins, including recombinant reverse transcriptase (rRT). These phosphorylations are selectively inhibited by CK-II inhibitors, such as quercetin, oGA (a glycyrrhetinic acid derivative) and NCS-chrom (an enediyne containing antibiotic). The data presented here suggest that HIV-1 Rev acts as an effective potent activator of CK-II, which may be a cellular mediator promoting HIV-1 replication in virus-infected cells.
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PMID:Biochemical characterization of HIV-1 Rev as a potent activator of casein kinase II in vitro. 965 40

DNA methylation of tumor suppressor genes is a common feature of human cancer. The cyclin-dependent kinase inhibitor gene p16/Ink4A is hypermethylated in a wide range of malignant tissues and the p14/ARF gene located 20 kb upstream on chromosome 9p21 is also methylated in carcinomas. p14/ARF (ARF, alternative reading frame) does not inhibit the activities of cyclins or cyclin-dependent kinase complexes; however, the importance of the two gene products in the etiology of cancer resides in their involvement in two major cell cycle regulatory pathways: p53 and the retinoblastoma protein, Rb, respectively. Distinct first exons driven from separate promoters are spliced onto the common exons 2 and 3 and the resulting proteins are translated in different reading frames. Both genes are expressed in normal cells but can be alternatively or coordinately silenced when their CpG islands are hypermethylated. Herein, we examined the presence of methyl-CpG binding proteins associated with aberrantly methylated promoters, the distribution of acetylated histones H3 and H4 by chromatin immunoprecipitation assays, and the effect of chemical treatment with 5-aza-2'-deoxycytidine (5aza-dC) and trichostatin A on gene induction in colon cell lines by quantitative reverse transcriptase-PCR. We observed that the methyl-CpG binding protein MBD2 is targeted to methylated regulatory regions and excludes the acetylated histones H3 and H4, resulting in a localized inactive chromatin configuration. When methylated, the genes can be induced by 5aza-dC but the combined action of 5aza-dC and trichostatin A results in robust gene expression. Thus, methyl-CpG binding proteins and histone deacetylases appear to cooperate in vivo, with a dominant effect of DNA methylation toward histone acetylation, and repress expression of tumor suppressor genes hypermethylated in cancers.
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PMID:Selective association of the methyl-CpG binding protein MBD2 with the silent p14/p16 locus in human neoplasia. 1130 12

The present study investigated the alteration of protein kinase C (PKC) isoforms in rat liver during the progression of sepsis. Cecal ligation and puncture (CLP) model of polymicrobial sepsis was used, with early and late sepsis referring to those animals sacrificed at 9 and 18 h, respectively, after CLP. The protein contents of various PKC isoforms were quantified by Western blot and densitometric analysis. PKCalpha activity was performed after immunoprecipitation and assayed based on the incorporation rate of 32p from [gamma-32p] adenosine triphosphate (ATP) into histone. The distribution of PKCalpha was evaluated by immunohistochemical staining. The steady state expression of PKCalpha mRNA was estimated by reverse transcriptase-polymerase chain reaction (RT-PCR). The results indicated that 1) five isoforms (alpha, beta, delta, epsilon, zeta) could be detected in normal rat liver. PKCalpha and beta were predominantly present in the cytosolic fraction, while membrane-associated PKCdelta was more prominent than that of cytosolic fraction; 2) the protein content of membrane-associated PKCalpha was significantly decreased at early (P < 0.05) and late (P < 0.01) sepsis; 3) there was no significant difference of protein contents of PKC-delta, -epsilon and -zeta between sham-operated and septic rat liver; 4) the activity of membrane-associated PKCalpha was significantly declined under detection level during sepsis; 5) at both early and late sepsis, the immunohistochemical staining of PKCalpha was significantly diminished, especially in the nucleus; 6) the RT-PCR product of PKCalpha mRNA of septic liver was significantly less than the sham-operated liver. These results suggest that inactivation and the suppression of PKC-alpha gene transcription might be involved in modulating hepatic failure during sepsis.
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PMID:Alteration of protein kinase C isoforms in the liver of septic rat. 1179 68

The localization of reverse transcriptase-related proteins in polytene chromosomes of dipterans was investigated using previously characterized antibodies to a recombinant polypeptide containing conserved motifs of insect reverse transcriptases. The immunoreactions were carried out with polytene chromosome squashes of eight sciarids, one chironomid and three Drosophila species. Telomeric staining was regularly observed on chromosomes of the sciarid Rhynchosciara americana under normal growth conditions. Five of eight chromosomal tips were labelled except for the heterochromatic ends that are occasionally found associated forming a chromocentre in the salivary gland. Reverse transcriptase-related proteins were detected at chromosomal tips of young larvae and remained bound to the telomeres throughout larval development. As in salivary gland chromosomes, five non-telocentric ends of the chromosomes from Malpighian tubules of R. americana appeared clearly stained with anti-reverse transcriptase. The occurrence of telomeric reverse transcriptase in R. americana correlates with the presence of RNA in addition to an unusual enrichment with homopolymeric dA/dT DNA associated with the telomeric heterochromatin. The antibodies also reacted with a few interstitial sites in chromosomes of four Rhynchosciara species, one band overlapping the histone gene locus of three species in the americana -like group. The results provide evidence for a reverse transcriptase-related protein as a constitutive component in telomeres of R. americana and also in certain interstitial loci of Rhynchosciara species in which RNA was immunologically detected in the form of RNA:DNA hybrids.
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PMID:Reverse transcriptase-related proteins in telomeres and in certain chromosomal loci of Rhynchosciara (Diptera: Sciaridae). 1270 82

Targeted expression of a human pituitary tumor derived-fibroblast growth factor receptor-4 (FGFR4) recapitulates pituitary tumorigenesis. We have shown that FGFR4 is a target for Ikaros, a zinc finger-containing transcription factor that localizes to heterochromatin regions and participates in higher order chromatin complexes and control of gene expression. We report here the expression of Ikaros and functional differences between its alternatively spliced variants in human pituitary tumors. Ik1 expression was detected in human pituitary tumors and we also identified a truncated isoform consistent with the non-DNA-binding Ik6 isoform in a subset of adenomas by reverse transcriptase-polymerase chain reaction, sequencing, and Western immunoblotting. Transfection of Ik6 in GH4 pituitary cells resulted in predominantly cytoplasmic expression as compared to Ik1, which resulted in exclusively nuclear expression as determined by immunofluorescence and immunoblotting of fractionated protein. Immunohistochemistry of primary human pituitary adenomas localized Ikaros expression to the nuclear compartment but also in the cytoplasm, the latter consistent with Ik6. Expression of Ikaros and truncated non-DNA-binding isoforms was also suggested by electromobility shift assays using nuclear proteins from primary human pituitary adenomas. Ik6 resulted in reversal of the effects of Ik1 on wild-type 5' FGFR4 promoter activity, histone acetylation, and regulation of the endogenous gene. We conclude that dominant-negative Ik6 isoforms with their distinct localization and effects on Ik1 action may contribute to the altered expression of FGFR4 and possibly other target genes in human pituitary tumors.
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PMID:Ikaros isoforms in human pituitary tumors: distinct localization, histone acetylation, and activation of the 5' fibroblast growth factor receptor-4 promoter. 2621 88


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