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)

The free 4S RNA of avian RNA tumor viruses is greatly enriched in one of the four methionine tRNAs of the host cells, tRNA4Met. On the assumption that viral tRNAMet forms are identical to the corresponding tRNAs of mouse or chick cells, the following conclusions were drawn concerning the tRNAMet content of oncornaviruses: (1) tRNAMet species may be compartmentalised within the host cells, and the viral tRNA pool could reflect the cellular compartment in which viral maturation takes place since tRNAMet forms distribute unevenly between different fractions of a cell homogenate. (2) tRNA4Met appears to have no special role in the modulation of protein synthesis in as much as no functional difference between tRNA2Met and tRNA3Met, tRNA4Met could be demonstrated in in vitro protein synthesising systems. (3) tRNA4Met differs in nucleotide sequence from all other host cell tRNAMet forms except possibly tRNA2Met. The nucleotide sequences of two tRNAMet species, tRNA1Met and tRNA4Met, have already been determined and the sequence of another host cell tRNAMet, tRNA3Met, was derived from the analogy of its sequence to that of tRNA4Met since the two molecules differ in only 6 nucleotides out of 76. (4) Avian myeloblastosis virus reverse transcriptase has been shown to bind specifically tRNA4Met and tRNATrp in whole cell tRNA and therefore the free tRNA4Met in the virion particle may exist substantially bound to virion-associated transcriptase.
Nucleic Acids Res 1978 Dec
PMID:Selection of methionine tRNAs by avian oncornaviruses. 21 69

The mRNA coding for the small apo-Very Low Density Lipoprotein (apo-VLDLII) from chicken serum was highly enriched by oligo(dT) chromatography and preparative gel electrophoresis of estrogenised liver RNA. Double-stranded cDNA was synthesised by the subsequent actions of reverse transcriptase and DNA polymerase, and used for a preliminary characterisation of the structural gene. Molecular cloning of dC-tailed ds-cDNA into the Pst I site of plasmid pBR 322 yielded several recombinant clones. Five chimeric DNAs were selected and characterised by restriction enzyme mapping and electron microscopy of R-loops. At least two of them (pVLDLII 3.33 and pVLDLII 4.82) contain an almost full-length ds-transcript of VLDLII mRNA in which no more than 10-20 bases at the 5'- end are missing.
Nucleic Acids Res 1979 Dec 20
PMID:Purification of the mRNA for chicken very low density lipoproteinII and molecular cloning of its full-length double-stranded cDNA. 23 Apr 63

A recombinant plasmid containing chick pro-alpha2 collagen gene sequences has been constructed and cloned in Escherichia coli. Using partially purified collagen mRNA as template, we synthesized double-stranded DNA by the successive action of reverse transcriptase (RNA-directed DNA nucleotidyltransferase) from avian myeloblastosis virus and the Klenow A fragment of E. coli DNA polymerase I. From this complex mixture of double-stranded DNAs, a specific 200-base-pair restriction fragment was generated by cleavage with the restriction endonucleases BamHI and EcoRI. These enzymes also make unique cuts in the plasmid vector pBR322. The restriction fragment was inserted into pBR322 via these BamHI and EcoRI sites and cloned in E. coli chi1776. The cloned recombinant plasmid was shown to contain pro-alpha2 collagen DNA by its specific hybridization to chick pro-alpha2 collagen mRNA, as assayed in an in vitro translation system. Thus, a clone containing pro-alpha2 collagen DNA was constructed without first obtaining highly purified collagen mRNA.
Proc Natl Acad Sci U S A 1978 Dec
PMID:Construction of a recombinant bacterial plasmid containing a chick pro-alpha2 collagen gene sequence. 36 5

Zein messenger RNAs from maize endosperm were purified by successive oligo(dT)-cellulose chromatography and sucrose gradient centrifugation. Polyacrylamide gel electrophoresis under denaturing conditions revealed the presence of two size classes of zein messenger RNAs of Mr 3.5 x 10(5) and 4.10 x 10(5). The mRNA was shown to synthesize the major zein polypeptides, to have a base composition characteristic of a poly(A)-containing RNA and to be transcribed by reverse transcriptase into complementary DNA. The r0t1/2 of the hybridization curve of cDNA hybridized to an excess of mRNA was shown to be 7 x 10(-2) M . s indicating that about 15 non-cross-hybridizing sequences are present in the zein mRNA preparations. The kinetics of cDNA annealing with an excess of maize DNA from 2 n cells suggest a ten-times reiteration of each mRNA sequence. This result is confirmed from saturation experiments, where in cDNA excess to DNA, the number of zein genes per haploid maize genome was estimated as about 120 copies. Similar experiments carried out on DNA from normal and mutant endosperms (3n cells) indicate the absence of large amplifications or deletions of zein genes in the tissue devoted to zein synthesis.
Eur J Biochem 1979 Dec
PMID:Genes and mRNAs coding for zein polypeptides in Zea mays. 52 Mar 23

A 226-nucleotide fragment was derived from alfalfa mosaic virus RNA 4 (ALMV RNA 4), the subgenomic messenger for viral coat protein, and its sequence was deduced by in vitro labeling with polynucleotide kinase and application of RNA sequencing techniques. The fragment contains the 3'-terminal 45 nucleotides of the coat protein cistron and the complete 3'-noncoding region of 182 nucleotides. The total length of RNA 4 was calculated to be 881 nucleotides. AlMV RNAs 1, 2 and 3 were elongated with a 3'-terminal poly(A) stretch and subjected to sequence analysis by using a specific primer, reverse transcriptase and chain terminators. This revealed and extensive homology between the 3'-terminal 140 to 150 nucleotides of all four ALMV RNAs. Despite a number of base substitutions, the secondary structure of the homologous region is highly conserved. The observed homology indicates that, as with RNA 4, the sites with a high affinity for the viral coat protein are located at the 3'-termini of the genomic RNAs.
Nucleic Acids Res 1979 Dec 11
PMID:Nucleotide sequence of the 3'-noncoding region of alfalfa mosaic virus RNA 4 and its homology with the genomic RNAs. 53 14

Synovial fluid lymphocytes from patients with rheumatoid arthritis have been examined for evidence of a productive infection with retroviruses by electron microscopy, labelling with 3H-uredine, growth in soft agar, and culturing in conditioned medium. No such viruses were detected. In addition, the synovial lymphocytes were activated before fusion and cocultivation with several cell lines which have proved permissive for primate retroviruses. Monitoring these cultures subsequently by reverse transcriptase assay, labelling with 3H-uridine, and membrane immunofluorescence gave no indication that retroviruses were present.
Ann Rheum Dis 1979 Dec
PMID:Viruses and lymphocytes in rheumatoid arthritis. I. Studies on cultured rheumatoid lymphocytes. 53 43

The rational design of antitumor and antiviral agents must ultimately take advantage of biochemical differences between normal host cells and transformed cells. The initial experiments must be performed with subcellular or cellular model systems. For the studies with arabinosyl nucleosides we have chosen those enzyme systems, synthesizing DNA and RNA; being precursor analogues, the different arabinosyl nucleosides have been added in the triphosphate state to the different DNA- and RNA polymerase assays. 1-beta-D-Arabinofuranosylcytosine-5'-triphosphate has been found to inhibit the RNA-dependent DNA polymerases (isolated from oncogenic RNA viruses) 200-fold more sensitively than viral and cellular DNA-dependent DNA polymerases. Recent results, showing that RNA-leukemia-virus-related sequences are present in DNA of some human leukemia patients might support the assumption that the efficacy of this antimetabolite in the treatment of acute leukemia is due to its, at least relative selective inhibitory activity on reverse transcriptase. 9-beta-D-Arabinofuranosyladenine-5'-triphosphate is a strong inhibitor of cellular DNA polymerases with the cytological consequence of an inhibition of cell proliferation. The clinical benefit of the compound in treatment of tumors is dependent on their levels of adenosine deaminase. The triphosphate of this compound is a 100-fold more sensitive inhibitor of the herpesvirus DNA polymerase compared to the cellular replicative DNA polymerase. In addition the analogue, incorporated into herpesvirus DNA, acts as chain terminator. These effects are the biochemical basis for the highly selective antiherpesvirus activity of this antimetabolite. The anomer 9-alpha-D-arabinofuranosyladenine-5'-triphosphate only inhibits cellular replicative DNA polymerase and has no effect on herpesvirus DNA polymerase. Consequently this agent acts only cytostatically and not antivirally. Concerning 1-beta-D-arabinofuranosyluracil and 1-beta-D-arabinofuranosylthymine no pronounced antitumor or antiviral effect is known.
Jpn J Antibiot 1977 Dec
PMID:Rational design of arabinosyl nucleosides as antitumor and antiviral agents. 61 2

Translation of Rauscher murine leukemia virus (R-MuLV) 35S RNA in an mRNA-dependent cell-free protein-synthesizing system yields polypeptides identical to authentic Pr65gag, the R-MuLV gag precursor, and Pr200gag-pol, the precursor to the R-MuLV reverse transcriptase. In addition to these polypeptides, the cell-free product contains a family of polypeptides of less than 65,000 molecular weight which appear to be generated by premature termination of protein synthesis within the viral gag gene. We compared the tryptic maps of several of these less than 65,000-molecular-weight premature termination polypeptides with that of full-size Pr65gag and found a progressive loss of tryptic peptides which could be assigned to known R-MuLV gag proteins. A 40,000-molecular-weight fragment, P40gag, lacked p10 and part of p30, placing p10 at the C terminus pf Pr65gag and p30 ajacent to it. Fragments of 33,000 (P33gag) and 27,000 to 28,000 (P27/28gag) molecular weight showed a successive loss of additional p30 tryptic peptides, but no loss of either p15 or p12. An 18,000-molecular-weight fragment lost p12 but retained p15. These data suggest an R-MuLV gag gene order of NH2-p15-p12-p30-p10-COOH.
J Virol 1978 Dec
PMID:Tryptic peptide analyses of polypeptides generated by premature termination of cell-free protein synthesis allow a determination of the Rauscher leukemia virus gag gene order. 73 99

Mitochondrial DNA polymerase was purified 2300-fold over isolated mitochondria from rat liver. Template-primer specificities of this enzyme were investigated. Activated DNA was satisfactorily used as an active template-primer, but both native and denatured DNAs showed a slight activity. Synthetic polynucleotide, poly(dA) - oligo(dT)10 was found to have a high efficiency under the same condition for activated DNA. When the closed-circular, nicked and gapped Co1E1 DNAs were employed as a template-primer, the enzyme could only utilize the gapped DNA, indicating that the displacement synthesis was not catalyzed by the enzyme itself. The enzyme also copied poly(A) - oligo(dT)10 in high efficiency at pH 7.5 in the presence of MnCl2. Such RNA-directed DNA polymerase activity of the enzyme was further characterized. Cofractionated endouclease activity was completely separated from the enzyme by glycerol gradient centrifugation.
Biochim Biophys Acta 1977 Dec 02
PMID:Template specificity of rat mitochondrial DNA polymerase. 92

Tests for the presence of oncornavirus-like particles in human biopsies were made by the Spiegelman simultaneous assay for 70S RNA and RNA-dependent DNA polymerase and by detection of 600-900S particles, incorporating 3H-uridine, produced by cultured biopsy cells. Thirty-one malignant melanoma biopsies from 29 patients were studied. Using the simultaneous assay, evidence of virus-like particles was found in 15/26 (58%) of melanoma biopsies, 0/3 naevi pools, 1/4 samples of skin adjacent to melanoma, 0/3 samples of normal adult skin and 0/3 prepuces. The velocity sedimentation technique was shown to be a useful screening test for oncornaviruses in studies of two virus-producing mouse cell lines (TKL-5 and WEHI-22), and was positive with 7/9 melanoma biopsies. Overall, these results are compatible with the earlier findings of similar virus-like particles in malignant melanoma cell lines, but the exact nature of the particles remains to be defined.
Int J Cancer 1976 Dec 15
PMID:Oncornavirus-like particles in malignant melanoma and control biopsies. 99 6


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