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)

Phosphonoacetate is a highly specific inhibitor of herpes simplex virus-induced DNA polymerase. Sensitivity of herpesvirus type 1 or type 2 induced DNA polymerase to the drug was similar. However, DNA polymerases from other sources such as the host cells (Wi-38), Micrococcus luteus, and hepatitis B virus were highly resistant. In addition, Escherichia coli RNA polymerase and reverse transcriptase of Rous sarcoma virus were also insensitive to the drug. Enzyme kinetic studies showed that inhibition was noncompetitive with respect to deoxyribonucleotide triphosphates. The Ki value was about 0.45 muM. The apparent Km values for dTTP, dATP, dCTP, and dGTP were 0.71, 0.75, 0.42, and 0.39 muM, respectively. The base composition of template has no profound effect on the extent of inhibition. The drug caused uncompetititve inhibition with respect to template which indicated that phosphonoacetate did not bind directly to template DNA. Results are presented which suggest that phosphonoacetate did not affect the formation of the enzyme-DNA complex but probably inhibited the elongation step of DNA polymerase reaction.
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PMID:Mode of inhibition of herpes simplex virus DNA polymerase by phosphonoacetate. 5 71

Three temperature-sensitive mutants of the Rauscher strain of murine leukemia virus are defective in early post-penetration functions required both for leukemia virus infection and for initiation of transformation of cells by their pseudotypes of murine sarcoma virus. In the present study, the reverse transcriptase of one of these mutants (ts 29) is shown to be thermolabile compared with the enzymes of the wild-type virus and several other temperature-sensitive mutants. These findings provide evidence that the reverse transcriptase is required both for leukemia virus infection and for initition of transformation by the replication-defective murine sarcoma virus genome.
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PMID:Thermolabile reverse transcriptase of a mammalian leukemia virus mutant temperature sensitive in its replication and sarcoma virus helper functions. 5 94

A cytoplasmic particulate fraction from human leukemic cells has been shown to contain reverse transcriptase and its associated high-molecular weight RHA template. We attempted to detect the reverse-transcriptase-template complex in morphologically normal peripheral blood leukocytes from patients with acute leukemia in complete remission. Our assay system consisted of a velocity glycerol gradient and cesium sulfate equilibrium gradient analysis of the endogenous reverse transcriptase reaction product. Three of nine patients in remission had positive reactions determined by glycerol gradient analysis, and eight of 10 patients in remission had positive reactions by cesium sulfate gradient analysis. We were unable to detect the template complex in leukocytes of normal persons. Thus, normal-appearing leukocytes in the peripheral blood of some leukemia patients in remission seem to retain a number of biochemical characteristics, possibly viral related, associated with leukemic cells.
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PMID:Reverse transcriptase in leukocytes of leukemic patients in remission. 5 87

Particles possessing a density of 1.16 g/ml and encapsulating a 70S RNA and a RNA-instructed DNA polymerase (reverse transcriptase) have been prepared from the spleen of a patient with chronic lymphocytic leukemia. These particles have been converted to cores with a density of 1.26 g/ml and containing the enzyme-RNA complex, in complete analogy to the known RNA tumor viruses of avian and murine origin. The reverse transcriptase was purified from the cores by column chromatography to a stage showing a single major protein band of 70,000 daltons in a gel electrophoresis. The enzyme was capable of transcribing heteropolymeric RNA into DNA complements as demonstrated by specific back hybridization to template RNA. The leukemic spleen would appear to represent an important source of this enzyme, as well as other potentially important leukemia-specific reagents.
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PMID:Purification of RNA-instructed DNA polymerase from human leukemic spleens. 5 34

Type-C RNA tumor virus particles were released from three different human lymphoblastoid cell lines after incubation in arginine-deficient medium. The released virus-like particles were characterized by (a) their ability to band in sucrose gradients at a density of 1.16-1.18 g/ml; (b) the presence of an RNA-directed DNA polymerase activity resembling that of the oncornaviruses; and (c) isolation of cores that band at a density of 1.26-1.27 g/ml in sucrose gradients. Examination of the arginine-deprived human lymphoblastoid cell line strain P3HR-1 by electron microscopy revealed the presence of C-type particles in the intracellular spaces.
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PMID:Oncornavirus-like particles released from arginine-deprived human lymphoblastoid cell lines. 5 42

Lens messenger ribonucleoprotein complexes have been isolated from calf lens polysomes by sucrose gradient centrifugation after puromycin-induced dissociation. A 10 S mRNA was released from a 13 S messenger ribonucleoprotein complex and a 14 S mRNA from a 19 S messenger ribonucleoprotein complex. Two major protein components with molecular weights of approx. 64 000 and 40 000 were isolated from each of the messenger ribonucleoprotein complexes after RNAase digestion. Buoyant density determinations suggest that the messenger ribonucleoprotein complexes contain approximately one mol of each major protein species per mol mRNA. In contrast to lens mRNA, lens messenger ribonucleoproteins are poor templates for transcription with avian myeloblastosis virus reverse transcriptase. Similar results were also obtained with globin messenger ribonucleoprotein containing either two major protein species (or deficient in the lower molecular weight protein species). Polynucleotide phosphorylase eliminates the reverse transcription template activity of the lens mRNA. This effect is blocked in the messenger ribonucleoprotein. Such observations suggest that at least one of the protein components associated with lens messenger ribonucleoprotein may be located in the 3'-terminal region. Only a small variation in translation activity was observed between the messenger ribonucleoproteins and their respective mRNAs.
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PMID:Calf lens messenger ribonucleoprotein complexes. Characterization and comparison of template activity with corresponding mRNAs. 5 93

A RNA-dependent DNA polymerase was isolated from a human cell line derived from the bone marrow of a patient with polycythemia vera. The purification procedure included chromatography on phosphocellulose and oligo(dT)-cellulose, and glycerol gradient centrifugation. The enzyme could be distinguished from polymerase A by salt elution from phosphocellulose, utilization of poly(rC) - oligo(dG) and its molecular size of about 70000, as determined by centrifugation. Throughout the purification procedure ribonuclease H activity was co-purified. Upon dodecylsulfate-polyacrylamide electrophoresis on microgradient gels two main bands with molecular weights of 68000 and 66000 and three minor bands were detected. The enzyme preferentially used poly(rA) - oligo(dT) as template-primer compared with poly(dA) - oligo(dT). It incorporated dGMP into polymer on poly(rC) - oligo(dG).
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PMID:RNA-dependent DNA polymerase from a cell line derived from the bone marrow of a patient with polycythemia vera. 5 57

A virus designated bovine leukaemia virus (BLV), associated with leukaemia in cattle and previously demonstrated to induce the disease in sheep, was purified from chronically infected sheep cell cultures. Electrophoretic analysis showed a major protein of mol. wt. about 24,000 (p24) which reacted in gel diffusion and complement-fixation tests with sera from naturally infected cattle, experimentally infected sheep, and guinea pigs immunized with p24. BLV p24 has an isoelectric point of 8-6. Interspecies antigenic reactivities characteristic of mammalian Type C virus p30s were not detected in disrupted BLV or on p24. Sheep and guinea pig antisera to BLV, reactive with p24, also did not precipitate several Type C virus p30s in radioimmunoassays. BLV is also distinguished from Type C viruses and resembles mouse mammary tumour virus and Mason-Pfezer virus in having an RNA-dependent DNA polymerase which is preferentially active in the presence of Mg++ when synthetic templates are used. Along with previously published morphological data, the above indicates that BLV is not a Type C virus as classically defined. Four hundred and forty one human sera from cancer patients and matched controls were non-reactive with disruped BLV, BLV infected cells, and BLV p24 in complement-fixation tests.
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PMID:Characteristics of the major internal protein and RNA-dependent DNA polymerase of bovine leukaemia virus. 5 5

A preliminary analysis of an RNA-directed DNA polymerase was made and a C-type virus-like particle was identified in platelets from 2 patients with the myeloproliferative disorder thrombocythemia (primary, essential, hemorrhagic, or idiopathic thrombocythemia). Platelet homogenates were centrifuged through a sucrose equilibrium density gradient. Both endogenous and exogenous DNA polymerase activity was found at a density of 1.19 g/ml. No activity was seen at comparable densities in control gradients. Electron micrographs of thin sections of these platelets revealed a particle with the morphologic characteristics of a C-type virus; however, the diameter of this particle was about 80 nm, slightly lower than that commonly found for C-type particles. Critical-point dried specimens, from the fractions of the sucrose gradient at which DNA polymerase activity was found, contained particles of the same size and morphology as those in the thin sections.
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PMID:Analysis of platelets from patients with thrombocythemia for reverse transcriptase and virus-like particles. 5 32

T24C, a continuous cell line derived from the pooled thymic tissue of normal inbred OM rats, spontaneously produced type-C virus. The virus genome was expressed cyclically. The amount of RNA-dependent DNA polymerase (RDP) and the number of 1.14 g dense particles/ml fluctuated simultaneously with cultivation. The released virus, RPT24C, did not infect cell lines from the rat, mouse, dog, or human. T31, also a rat thymus line, during its 2.5 years of cultivation did not produce type-C virus. Cocultivation with potentially permissive lines did not rescue any virus. 5-lodo-2'-deoxyuridine treatments at earlier passages yielded negative results. Chemical treatment at passages 111, 116, 123, and 128 yielded varying amounts of 3H-uridine incorporation at a sucrose density of 1.14 g/ml. Enzyme assays on chemically treated T31 cultures tested at passage 111 showed a small but transient burst of RDP activity. T31-B, a subline of T31, which was frozen and thawed once, released rat type-C virus spontaneously at passage 56. Two additional sublines of T31 (NI-T31 and NII-T31) were maintained for 2.5 years in culture without any cell-dispersing treatment. NI-T31, but not NII-T31, spontaneously released type-C virus. Once induced, the type-C viruses from T31-B and NI-T31 were continuously produced.
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PMID:In vitro activation, infectivity, and production of endogenous type-C virus from OM rats. 5 37


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