Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

Association between mycosis fungoides (MF), its leukaemic variant Sezary syndrome (SS) and the human T-cell lymphotropic virus type-I (HTLV-I) has been controversial, with the reported incidence of infection varying between 0% and nearly 100%. We studied 127 patients (85 MF, 28 SS, five Sezary cell leukaemia, four lymphomatoid papulosis, and five unspecified cutaneous T-cell lymphomas (CTCL)) originating from Europe (France, Spain, U.K., Portugal) or from U.S.A. (California) for the presence of HTLV-I infection markers. HTLV-I and -II serology were performed on 78 patients using standard immunological methods. Reverse transcriptase (RT) assay was also performed in 26 cases using an RT-PCR-based method of high sensitivity. Molecular analyses were performed on 215 DNA samples (121 from fresh PBMCs, 26 from PBMCs after short-term culture and 68 from skin lesions) by PCR amplification using HTLV-I and -II gag, pol, env, pX and LTR specific primers. Immunological tests were negative except for two sera which were indeterminate. PCR with all HTLV-I and -II primer pairs showed negative results in all 215 samples investigated. No RT activity was detected in short-term PBMC cultures of any of the 26 cases studied. The results of this large study from five different countries clearly indicate that MF and SS are not associated with HTLV-I infection.
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PMID:Mycosis fungoides and Sezary syndrome are not associated with HTLV-I infection: an international study. 1055 37

The non-segmented, double-stranded RNA genome of the Giardia lamblia virus (GLV) contains two genes encoding the major capsid protein (gag) and a fusion of gag with the viral RNA-dependent RNA polymerase (pol). Computer analysis of the viral RNA genome revealed three putative stem-loop structures that were predicted to mediate replication, transcription and packaging of the GLV genomic RNA by binding to the pol domain of the virus-encoded fusion protein. To provide evidence of these postulated RNA/protein interactions, gel retardation assays were employed to examine the potential binding capacity of various viral RNA genome-related sequences to native GLV protein(s). Viral proteins were obtained by disrupting purified GLV particles under low-ionic-strength conditions. The resulting viral protein particles maintained their RNA polymerase activity in the presence of GLV genomic RNA and thus appeared to be suitable tools for the analyses of GLV-protein-mediated binding reactions. A 72-nt short single-stranded in vitro transcript containing a putative stem-loop structure predicted to participate in the packaging of GLV (+)-strand RNA bound specifically to the disrupted virus particles. RNAs containing modified motifs of this stem-loop structure failed to bind to the GLV capsid.
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PMID:Site-specific binding of polymerase-containing particles of the Giardia lamblia double-stranded RNA virus to the viral plus-strand RNA. 940 30

The nucleotide sequences of the genome RNA encoding the RNA polymerase and the 3' non-coding region (NCR) of bovine enterovirus (BEV) serotype I Japanese isolate, MZ468, were determined. The genetic distance between the two BEV serotype I strains, MZ468 and VG-5-27, was calculated by pairwise comparison of nucleotide sequences. The synonymous substitution rate was high (1.40 x 10(-2)/site/year), and of the same order as those of influenza virus HA, HIV-1 gag and env, and enterovirus 70 VP1 genes.
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PMID:Nucleotide sequence of the gene encoding the RNA polymerase and the 3' non-coding region of a bovine enterovirus Japanese isolate: rapid synonymous substitutions between European and Japanese strains. 963 51

A human cell line constitutively expressing the HIV-1 gag and pol genes products was established. The cell line was established by stably transfecting 293 cells with a plasmid construct that expresses the HIV Gag and Pol and can confer the transfectants resistant to mycophenolic acid. Particles generated from transient expression of the plasmid construct were noninfectious when pseudotyped with HIV envelope or with amphotropic murine leukemia virus envelope proteins. However, virus-like Gag particles produced by the stable cell line were appropriately processed, exhibited a wild-type retrovirus particle density, and possessed significant reverse-transcriptase (RT) activities. Continuous passage of the cell line either in the presence or absence of mycophenolic acid had no major effects on the Gag processing efficiency, particle assembly, or RT activity release. It was also demonstrated that the proteolytic processing of the virus-like particles released from the cell line was inhibited by an HIV protease inhibitor, saquinavir. The establishment of a stable cell line producing noninfectious but proteolytically processed HIV Gag particles offers a safe, convenient tool for biochemical and immunological analysis of virus-like particle assembly and is very useful for the development of anti-HIV protease drugs.
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PMID:A human cell line constitutively expressing HIV-1 Gag and Gag-Pol gene products. 989 Apr 17

Reverse transcriptase (RT) activity has been detected recently in all chicken cell-derived measles and mumps vaccines. A study of a vaccine manufactured in Europe indicated that the RT is associated with particles containing endogenous avian retrovirus (EAV-0) RNA and originates from the chicken embryonic fibroblasts (CEF) used as a substrate for propagation of the vaccine. We investigated the origin of RT in measles and mumps vaccines from a U.S. manufacturer and confirm the presence of RT and EAV RNA. Additionally, we provide new evidence for the presence of avian leukosis virus (ALV) in both CEF supernatants and vaccines. ALV pol sequences were first identified in particle-associated RNA by amplification with degenerate retroviral pol primers. ALV RNA sequences from both the gag and env regions were also detected. Analysis of hypervariable region 2 of env revealed a subgroup E sequence, an endogenous-type ALV. Both CEF- and vaccine-derived RT activity could be blocked by antibodies to ALV RT. Release of ALV-like virus particles from uninoculated CEF was also documented by electron microscopy. Nonetheless, infectivity studies on susceptible 15B1 chicken cells gave no evidence of infectious ALV, which is consistent with the phenotypes of the ev loci identified in the CEF. PCR analysis of ALV and EAV proviral sequences in peripheral blood mononuclear cells from 33 children after measles and mumps vaccination yielded negative results. Our data indicate that the sources of RT activity in all RT-positive measles and mumps vaccines may not be similar and depend on the particular endogenous retroviral loci present in the chicken cell substrate used. The present data do not support transmission of either ALV or EAV to recipients of the U.S.-made vaccine and provide reassurance for current immunization policies.
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PMID:Evidence of avian leukosis virus subgroup E and endogenous avian virus in measles and mumps vaccines derived from chicken cells: investigation of transmission to vaccine recipients. 1036 36

CD40 ligand (CD40L), expressed on activated T cells, binds its receptor, CD40, on dendritic cells, B cells, and monocytes/ macrophages. Human immunodeficiency virus (HIV)-infected individuals exhibit normal B-cell CD40 expression but diminished expression of CD40L on CD4 + T cells. Thus, we studied recombinant human CD40L (huCD40L) in an in vitro rhesus macaque model of acquired immunodeficiency syndrome (AIDS). huCD40L induced peripheral blood mononuclear cell (PBMC) proliferation independent of mitogenic cytokines and led to a 70% reduction in p27 production by simian immunodeficiency virus (SIV) mac239 infected PBMCs (P < 0.05). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed reduced expression of SIV gag and increased expression of interleukin (IL)-16 mRNA. Supernatants from huCD40L-stimulated PBMC and control cultures contained similar amounts of IL-16, suggesting an intracellular antiviral effect by IL-16. Phytohemagglutinin (PHA)-stimulated PBMCs similarly cultured with huCD40L showed only slight increases in chemokine production (P > 0.05). These results suggest that huCD40L inhibits replication (antigen and mRNA production) of SIVmac239. This response involves huCD40L induction of IL16 mRNA expression and appears to be independent of beta-chemokines.
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PMID:Recombinant human CD40 ligand inhibits simian immunodeficiency virus replication: a role for interleukin- 16. 1059 85

Pigs are potential providers of donor tissues for xenotransplantation (e.g. of pancreatic islets) in Type 1 diabetes. In this context, our group has studied the use of islets from specific pathogen-free (SPF) pigs as a means of reducing the risks of "conventional zoonosis". Although this approach does not prevent the transmission of pig endogenous retrovirus (PERV) to humans, we attempted to determine the presence of C-type PERV mRNAs for gag, pol, and env subtypes as a first descriptive step in the retroviral characterisation of SPF pig tissues (especially pancreas). Using semiquantitative reverse-transcriptase polymer chain reaction with 18S rRNA and beta-actin as internal controls, PERV mRNA levels were compared in a large panel of tissues from SPF and conventional pigs. PERV mRNAs for gag, pol, env-A and env-B were present in all tissues studied from the nine SPF pigs tested. Signals for env-C mRNAs were of much lower intensity than those for env-A and B, and most often undetectable in pancreas. The mRNA levels for gag, pol, env-A, env-B and env-C mRNAs were lower in pancreas (p < 0.01) than in all other tissues. Among other porcine tissues likely to be grafted in man, the highest retroviral mRNA levels were detected in kidney (p < 0.01), followed by liver, lung and heart. Amplified PERV mRNA signals were about 17 times less frequent in pig pancreas than in the retroviral-producing porcine cell line G2, while kidney contained about 6 times more PERV mRNAs than pancreas. The levels of gag, pol, env-A, env-B, and env-C mRNAs also varied between tissues of conventional pigs: PERV mRNA levels were lowest in pancreas, and env-C mRNAs were most often undetectable. For all SPF tissues tested, pol, gag, env-A, env-B, and env-C mRNA levels were in the same range or slightly higher than in corresponding tissues of conventional pigs. In summary, this study of C-type PERV mRNAs in a large panel of tissues from SPF pigs, in the context of our strategy of quality assurance and sanitary control, indicated that PERV mRNA levels were in the same range in SPF and corresponding conventional pig tissues, confirming that the use of SPF pigs would not prevent the risk of PERV transmission to human recipients of xenografts. PERV-A and PERV-B may be mainly represented, and PERV-C much less, in these pig tissues (particularly pancreas). The fact that pancreas expressed the lowest PERV mRNA levels and kidney the highest, among porcine tissues likely to be grafted, could be of interest from a clinical point of view. Pig tissues may differ in their loads of PERV sequences, which could be a factor in the risk of PERV transmission during xenotransplantation.
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PMID:Porcine endogenous retroviral mRNAs in pancreas and a panel of tissues from specific pathogen-free pigs. 1063 79

Since 1984, unheated porcine clotting factor VIII (Hyate:C) has been used to treat severe bleeding episodes in persons with hemophilia who have antibodies to human clotting factor. We document the presence of porcine endogenous retrovirus (PERV) in plasma samples of pigs and in clinical lots of Hyate:C. Both gag and pol PERV RNA sequences were detected by reverse-transcriptase (RT) polymerase chain reaction in 13 of 13 lots of Hyate:C tested. Among 10 of these lots, RT activity also was detected, which confirms the presence of retroviral particles. To assess the transmission of PERV to Hyate:C recipients, we tested serum specimens from 88 recipients of Hyate:C and 23 noninfused control subjects for anti-PERV antibodies by using a Western blot assay. None of the samples was positive. Our data document that PERV particles are a common contaminant of Hyate:C products and suggest that the risk of PERV transmission from these percutaneous exposures is very low.
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PMID:Evidence of porcine endogenous retroviruses in porcine factor VIII and evaluation of transmission to recipients with hemophilia. 1117 Sep 92

We characterized the consensus sequence and structure of a long terminal repeat (LTR) retrotransposon from the genome of the human blood fluke, Schistosoma japonicum, and have named this element, Gulliver. The full length, consensus Gulliver LTR retrotransposon was 4788 bp, and it was flanked at its 5'- and 3'-ends by LTRs of 259 bp. Each LTR included RNA polymerase II promoter sequences, a CAAT signal and a TATA box. Gulliver exhibited features characteristic of a functional LTR retrotransposon including two read through (termination) ORFs encoding retroviral gag and pol proteins of 312 and 1071 amino acid residues, respectively. The gag ORF encoded motifs conserved in nucleic acid binding proteins, while the pol ORF encoded conserved domains of aspartic protease, reverse transcriptase (RT), RNaseH and integrase, in that order, a pol pattern conserved in the gypsy lineage of LTR retrotransposons. Whereas the sequence and structure of Gulliver was similar to that of gypsy, phylogenetic analysis revealed that Gulliver did not group particularly closely with the gypsy family. Rather, its closest relatives were a LTR retrotransposon from Caenorhabditis elegans, mag from Bombyx mori and, to a lesser extent, easel from the salmon Oncorhynchus keta. Dot blot hybridizations indicated that Gulliver was present at between 100 and several thousand copies in the S. japonicum genome, and Southern hybridization analysis suggested its probable presence in the genome of Schistosoma mansoni. Transcripts encoding the RT domain of Gulliver were detected by RT-PCR in larval and adult stages of S. japonicum, indicating that (at least) the RT domain of Gulliver is transcribed. This is the first report of the sequence and structure of an LTR retrotransposon from any schistosome or indeed from any species belonging to the phylum Platyhelminthes.
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PMID:Gulliver, a long terminal repeat retrotransposon from the genome of the oriental blood fluke Schistosoma japonicum. 1124 79

The RNA polymerase of giardiavirus (GLV) is synthesized as a fusion protein through a -1 ribosomal frameshift in a region where gag and pol open reading frames (ORFs) overlap. A heptamer, CCCUUUA, and a potential pseudoknot found in the overlap were predicted to be required for the frameshift. A 68-nucleotide (nt) cDNA fragment containing these elements was inserted between the GLV 5' 631-nt cDNA and the out-of-frame luciferase gene that required a -1 frameshift within the 68-nt fragment for expression. Giardia lamblia trophozoites transfected with the transcript of this construct showed a frameshift frequency at 1.7%, coinciding with the polymerase-to-capsid protein ratio in GLV. The heptamer is required for the frameshift but can be replaced with other sequences of the same motif. Mutations placing stop codons in the 0 or -1 frame, located directly before or after the heptamer, implicated the latter as the site for the -1 frameshift. Shortening or destroying the putative stem decreased the frameshift efficiency threefold; the efficiency was fully recovered by mutations to restore the stem. Deleting 18 nt from the 3' end of the 68-nt fragment, which formed the second stem in the putative pseudoknot, had no effect on the frequency of the frameshift. Chemical probing of the RNA secondary structure in the frameshift region showed that bases resistant to chemical modification were clustered in the putative stem structures, thus confirming the presence of the postulated stem-loop, while all the bases in the loop were chemically modified, thus ruling out their capability of forming a pseudoknot. These results confirmed the conclusion based on data from the mutation study that there is but a simple stem-loop downstream from the heptamer. Together, they constitute the structural elements for a -1 ribosomal frameshift in the GLV transcript.
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PMID:Structural analysis of the -1 ribosomal frameshift elements in giardiavirus mRNA. 1160 3


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