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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)

RNA polymerase activity of nuclei and positive circular dichroism ellipticity in the 250--300 nm region of chromatin were studied in three lymphoblastoid cell lines: HRI, an Epstein-Barr Virus (EBV) positive virus-producing line; Raji, and EBV positive virus-producing line; Raji, an EBV positive non-producer line; and SU-AMB-1, and EBV negative line. Raji consistently exhibited the highest and SU-AMB-1 the lowest polymerase activity and ellipticity. All lines showed higher RNA polymerase activity and positive ellipticity when exponentially growing than when stationary. In Raji it was shown that changes in RNA polymerase activity and chromatin ellipiticity were parallel. The changes in RNA polymerase activity of nuclei and circular dichroism ellipticity correlated with those of the proliferative state but not with the number of viral genomes or the production of virus or viral antigens.
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PMID:Transcription and circular dichroism of chromatin in lymphoblastoid cell lines during proliferation and quiescence. 71 33

The genome of Epstein-Barr virus (EBV) codes for two non-translated small RNA molecules, EBER 1 and 2. We found that both EBERs are expressed in the major EBV-carrying cell types, group I and III Burkitt's lymphoma (BL) cell lines, lymphoblastoid cell lines (LCLs) and in two nude mouse-passaged nasopharyngeal carcinoma (NPC) tumours. The relative amount of EBER 1 and EBER 2 varied in different host cells but did not correlate with the cellular phenotype. The EBER coding and flanking sequences were predominantly hypomethylated at HpaII sites not only in LCLs which usually carry hypomethylated EBV genomes but also in BL and NPC cell lines harbouring EBV episomes that are highly methylated in other regions. Thus, the EBER transcription units, actively transcribed by RNA polymerase III in the major EBV-carrying cell types, represent a methylation-free region in the EBV genome similarly to regulatory sequences of the latent membrane protein gene when the latter is transcribed by RNA polymerase II.
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PMID:RNA polymerase III-transcribed EBER 1 and 2 transcription units are expressed and hypomethylated in the major Epstein-Barr virus-carrying cell types. 132 Dec 9

A clone of the Epstein-Barr virus (EBV) thymidine kinase (TK) gene was derived from a cDNA library of P3HR1 cells. The gene product was expressed as a fusion protein in a procaryotic system by using T7 RNA polymerase. The recombinant TK showed a molecular mass of 67 kDa and was biologically active. Antiserum raised in mice immunized with partially purified TK recognized an antigen present in EBV-superinfected Raji cells using an indirect immunofluorescence assay.
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PMID:Cloning and expression of a cDNA encoding the Epstein-Barr virus thymidine kinase gene. 133 Nov 57

Adeno-associated virus (AAV) is a nonpathogenic parvovirus which normally requires helper adenovirus or herpes-virus for replication. We examined the growth of AAV type 2 in human lymphocytes and its possible interaction with HIV-1. Three B cell lines (CK-B, HS-2, and UC729) and four T cell lines (Molt-4, Jurkat, HUT78, and HUT78+HIV, which is persistently infected with HIV-1) were infected with AAV either in the presence or in the absence of adenovirus. AAV DNA was found in cells of all the lines following incubation with the virus, indicating absorption. AAV DNA replication occurred in most cell lines without particular preference for B or T cells, but only in the presence of helper virus, either adenovirus or Epstein-Barr virus. Expression of AAV proteins was examined by immunoblotting and ELISA, using sera specific for AAV Rep or capsid proteins. The level of AAV protein synthesis correlated with the efficiency of AAV DNA replication, and both varied between cell lines. The yield of infectious AAV was low in most cases, except in one T4 line (Jurkat), where AAV replication and protein synthesis in the presence of adenovirus were very extensive. In HUT78+HIV cells both adenovirus and AAV (in the presence of Ad2) replicated efficiently. The effects of adenovirus plus AAV coinfections on HIV-1 replication, measured by reverse-transcriptase (RT) activity, were mild. Infection with adenovirus or AAV alone resulted in a 60-70% increase in RT activity, while infection with AAV plus adenovirus resulted in a 20% decrease in RT activity. The yield of infectious AAV in this cell line was very low.
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PMID:Replication of adeno-associated virus type 2 in human lymphocytic cells and interaction with HIV-1. 137 38

We have studied the relative rate of transcription across the Epstein-Barr virus genome in the Burkitt's lymphoma cell line Raji by nuclear run-on analysis during latency and after induction of an abortive lytic cycle with 12-0-tetradecanoylphorbol 13-acetate (TPA) and 5-iodo-2'-deoxyuridine (IUdR). During latency the entire, or almost the entire, viral genome was found to be transcriptionally active to a low or intermediate extent, with some variation in activity along the genome. The fragment with the highest transcriptional activity was EcoRI J, which contains the genes encoding the small nuclear RNAs EBER1 and -2, transcribed predominantly by RNA polymerase III. An intermediate level of transcription was observed between positions 10 and 138 (kb), with areas of slightly higher activity on the large internal repeats and the left duplicated region (DL). The remaining part of the viral genome, between position 138 and the termini, and the termini and position 10 (kb) (with the exception of the EcoRI J fragment), showed very little transcriptional activity, except for the intermediately active regions carrying the righthand oriLyt (DR) and the terminal repeats. Upon induction of the viral genome with TPA and IUdR, the viral genome was transcriptionally active at a rate at least tenfold that seen during latency. Polymerases were not equally distributed along the genome after induction; the highest density was found in regions 48 to 58 kb, 82 to 84 kb, 102 to 104 kb, 118 to 122 kb and 142 to 145 kb of the viral genome. High transcriptional activity correlated with distinct transcription units in some cases, i.e. BamHI H1LF1 (DL), BamHI MLF1, BamHI ZLF1/BamHI RLF1 and BamHI X (thymidine kinase), but not in others (BamHI H2). Besides initiation of transcription, other regulatory processes such as stabilization and processing of primary transcripts may also contribute to regulation of virus gene expression. Addition of cycloheximide completely abolished the transcriptional activation of the genome mediated by TPA and IUdR.
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PMID:Transcriptional activity across the Epstein-Barr virus genome in Raji cells during latency and after induction of an abortive lytic cycle. 165 54

We found that hydrolysates of poly(A)+ RNA from Ehrlich ascites carcinoma cells which were transcribed by RNA polymerase III contained an unusual component designated as X. It was part of B2 RNA representing a transcript of B2 retroposon, typical of rodents. The component X possesses a cap-like structure, xppp5'G, where x has a non-nucleotide structure. About half of all B2 RNAs contained this group at the 5' end. Previously, Epstein et al. (1) detected a similar structure at the 5' end of small nuclear U6 RNA. Later, Singh and Reddy (2) showed methyl to be the blocking group in the component x of U6 RNA. Besides B2 RNA, we found 5' ends containing methyl groups in 7SK RNA.
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PMID:B2 RNA and 7SK RNA, RNA polymerase III transcripts, have a cap-like structure at their 5' end. 170 Aug 54

We found that hydrolysates of poly(A)RNA from Ehrlich ascites carcinoma cells which were transcribed by RNA polymerase III contained an unusual component designated as X. It was part of B2 RNA representing a transcript of B2 retroposon, typical of rodents. The component X possesses a cap-like structure, xPPP5'G, where x has al non-nucleotide structure. About half of all B2 RNAs contained this group at the 5'-end. Previously, Epstein et al. (Epstein P., Reddy R., Henning D., Busch H. parallel J. Biol. Chem. 1980. V. 255. P. 8901-8906) detected a similar structure at the 5'-end of small nuclear U6RNA. Later, Singh and Reddy (Singh R., Reddy R. parallel Proc. Natl. Acad. Sci. USA. 1989. V. 86. P. 8280-8283) showed methyl to be the blocking group in the component X of U6RNA. Besides B2 RNA, we found 5'-ends containing methyl groups in 7S K-RNA.
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PMID:[B2 RNA and 7SK RNA, transcripts of RNA-polymerase III, have a cap-like structure at the 5'-end]. 171 20

Human B lymphocytes latently infected with Epstein-Barr virus (EBV) synthesize very large amounts (5 x 10(6)/cell) of two small nuclear RNAs called EBERs (Epstein-Barr encoded RNAs). These RNAs are of unknown function and, like many RNA polymerase III (Pol III) transcripts, bind the La autoantigen. We have discovered that the EBERs also associate with a second highly abundant host-encoded protein designated EAP (EBER associated protein). Human EAP is a small (14,777 dalton, 128 amino acid) polypeptide that binds both EBER 1 and EBER 2. EAP is also found in association with one or both of two analogous virally-encoded RNAs found in baboon cells infected with herpesvirus papio (HVP). We have devised a purification procedure for EAP and have cloned its cDNA from a human placental cDNA library using amino acid sequence data and the polymerase chain reaction (PCR). The predicted amino acid sequence of EAP shows a strong resemblance (77% identity) to an endodermal, developmentally regulated sea urchin protein called 217 (Dolecki et al., 1988). EAP contains a potential nuclear localization signal and a highly acidic carboxy terminus, but does not display marked similarity to any other RNA binding proteins.
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PMID:EAP, a highly conserved cellular protein associated with Epstein-Barr virus small RNAs (EBERs). 184 7

Nuclear run-on assays revealed extensive transcription of the Epstein-Barr virus genome during latent infection in in vitro-infected human fetal lymphoblastoid cells (IB-4). The EBER genes were the most heavily transcribed viral genes in these cells. Their transcription was partially inhibited in the presence of 1 microgram of alpha-amanitin per ml and fully inhibited at 100 micrograms/ml, consistent with RNA polymerase III transcription. All other transcription was inhibited at 1 microgram of alpha-amanitin per ml, consistent with RNA polymerase II sensitivity to alpha-amanitin. Other than EBER transcription, almost no transcription occurred from the U1 region. Specifically, no transcription was detected from the U1 latent promoter. RNA polymerase II transcription was highest in IR1, extending rightward through U2 and IR2 into the U3 domain and gradually decreased, but was measurable throughout the rest of the genome. This is consistent with EBNA gene transcription initiation within IR1. The higher level of transcription of the IR1 and U2 domains, which encode EBNA-LP and EBNA-2, as opposed to the domains which encode EBNA-3A, EBNA-3B, or EBNA-3C or EBNA-1, correlated with a higher level of EBNA-LP/EBNA-2 mRNA. Transcription extended through U4 into U5, even though no known latent-gene mRNAs are expressed from U4 downstream of the EBNA-1 open reading frame. This may result from inefficient termination of EBNA gene transcription. Leftward transcription from the latent membrane protein promoter was lower than EBNA transcription, although the latent membrane protein mRNA was the most abundant of the latent-gene mRNAs, indicating that this mRNA is more efficiently processed or has a longer half-life. Although transcription was detected from the DL strong early promoters and to a lesser extent from other early promoters, early mRNAs were less abundant than EBNA mRNAs or undetectable, suggesting that there may be posttranscriptional as well as transcriptional control over early mRNA expression in these latently infected cells.
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PMID:Transcription of the Epstein-Barr virus genome during latency in growth-transformed lymphocytes. 215 49

The Epstein-Barr virus-encoded small RNA (EBER) genes appear to comprise an interesting subset of class III genes different from any previously identified, including U6 and 7SK. EBER genes have functional A and B box intragenic control regions. In addition, they contain three upstream elements that together stimulate in vivo expression 50-fold and resemble sites associated with typical class II promoters. DNAase I footprinting analyses using purified proteins or oligonucleotide competition demonstrate that nucleotides -40 to -55 bind activating transcription factor (ATF) or a related protein, while nucleotides -56 to -77 bind Sp1 protein or a related protein. The element between positions -23 and -28 resembles a TATA box. EBERs are unusual RNA polymerase III transcripts shown to be controlled by ATF- and Sp1-like promoter elements, suggesting mechanisms for their high level expression in EBV-transformed lymphocytes.
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PMID:Epstein-Barr virus small RNA (EBER) genes: unique transcription units that combine RNA polymerase II and III promoter elements. 254 26


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