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)

A bacteriophage gamma Ch4A clone containing a 22-kb rat DNA insert was isolated and found to contain a solitary tRNA(Phe)GAA gene and, 436 bp downstream of it, an Alu-like element. The nucleotide sequence of a 1141-bp DNA fragment containing these genes was determined. The rat tRNA(Phe)GAA gene, with the exception of an additional A in the extra arm, has a sequence identical to that of a rabbit liver tRNA(Phe). The Alu-like element belongs to the rodent B2 family of short interspersed repetitive nucleotide sequences. This repetitive element, B2Phe, is flanked by 12-bp direct repeats, contains an internal split promoter (block A and block B) for RNA polymerase III and is devoid of an A-rich segment at the 3' end. Like other members of the B2 family, the B2Phe element presents 64% sequence homology with rat serine tRNA and contains a serine (GCT) anticodon. Both tRNA(Phe)GAA gene and B2Phe element were found to be transcriptionally active in HeLa cell and Xenopus oocyte nuclear extracts. The tRNA(Phe) gene transcripts were processed during the course of transcription to form mature-size tRNA(Phe). The transcription efficiency of the B2Phe element was found to be an order of magnitude higher than that of the tRNA(Phe) gene. Competition experiments demonstrate that the B2Phe DNA can form a more stable transcription complex than the tRNA(Phe) gene and compete with it for binding of transcription factors.
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PMID:Nucleotide sequence and transcription of a rat tRNA(Phe) gene and a neighboring Alu-like element. 323 68

The potent environmental carcinogen benzo[a]pyrene (BaP), following enzymatic activation to enantiomeric pairs of bay-region 7,8-diol 9, 10-epoxides (the benzylic 7-hydroxyl group and epoxide oxygen are cis for DE-1 diastereomers and trans for DE-2 diastereomers), reacts with DNA to form covalent adducts predominately at the exocyclic amino groups of purines. Specific adducts, corresponding to the trans opening of each of the four optically active BaP DE isomers at C-10 by the N 2-amino group of dG, were synthesized as appropriately blocked phosphoramidites and were incorporated at either the first or second G of codon 12 within the G-rich sequence of human K-ras codons 11-13: GCT G1G2T GGC. The adducted oligonucleotides were incorporated into plasmids by primer extension, followed by purification of the covalently closed circular constructs. Adducts derived from either (+)- or (-)-DE-2, placed at either G1 or G2, presented strong blocks to in vitro transcription elongation by bacteriophage T3 RNA polymerase, but only moderately blocked transcription elongation by human RNA polymerase II in nuclear extracts. Adducts derived from all four DEs, placed on either G1 or G2, were used as substrates in a DNA repair synthesis assay using human whole cell extracts. Adducts derived from three of the DE stereoisomers exhibited significant amounts of repair synthesis, but the (-)-DE-2 adduct experienced no repair synthesis above that of the control. Constructs containing a pre-existing nick at the sixth phosphodiester bond 3' to either (+)-DE-2 or (-)-DE-2 adducts exhibited increased repair synthesis.
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PMID:Stereospecific differences in repair by human cell extracts of synthesized oligonucleotides containing trans-opened 7,8,9, 10-tetrahydrobenzo[a]pyrene 7,8-diol 9,10-epoxide N2-dG adduct stereoisomers located within the human K-ras codon 12 sequence. 988 96

During the winter of 2000, tomatoes (Lycopersicon esculentum) with a bright yellow leaf mosaic were observed in a commercial greenhouse in southern Ontario, Canada. Examination of leaf extracts, using leaf dips and immunosorbent absorption electron microscopy (ISEM), showed flexuous rods consistent with the potexvirus group. Polyclonal antibodies raised against the original Peruvian Pepino mosaic virus (PepMV) isolate (1) and commercial antibodies obtained from Deutsche Sammlung von Mikro-organismen und Zellkulturen (DSMZ), GmbH, Braunsweig, Germany, and Plant Research International (PRI), Wageningen, the Netherlands, were used in ISEM. Leaves tested positive in double-antibody sandwich-enzyme-linked immunosorbent assay (ELISA) with antibodies from DSMZ and PRI. A triple-antibody sandwich-ELISA obtained from Adgen Ltd. (Nellies Gate, UK) gave similar results. Potato virus X did not react with PepMV antiserum in ELISA. Positive PepMV ELISA controls were a U.K. and a Dutch isolate supplied by R. Mumford and R. A. A. van Vlugt, respectively, and DSMZ. Using primers generated from a sequence of the RNA polymerase region of a U.K. PepMV isolate (R. Mumford, unpublished data), a reverse transcription-polymerase chain reaction test showed the expected 312-bp amplicon for the Canadian, Dutch, and U.K. isolates. The primer sequences used were forward 5' CTA TTA CAA CTC CGG AAG CCA 3' and reverse 5' TGG TCT GGC CAG GCT TTG AC 3'. The three isolates were maintained in tomato cv. Bush Beefsteak. When mechanically inoculated on L. esculentum cv. Rapsodie, the Canadian isolate caused a bright yellow mosaic in 1 to 2 weeks, while the two European isolates caused a faint yellow mosaic and mild puckering of the leaves. When mechanically inoculated on 17 indicator plants, the Canadian isolate had a host range similar to the U.K. isolate. The most striking difference in symptoms occurred in L. pimpinellifolium, in which the Canadian isolate caused a yellow mosaic, the Dutch isolate caused no symptoms, and the U.K. isolate caused a marked puckering of the leaves, suggesting virus strain differences among the isolates. Tomato fruits originating from the United States were collected during border inspections by the Canadian Food Inspection Agency and tested for PepMV by ELISA with antisera from DSMZ. PepMV was not detected in 7 samples from California, but was detected in 6 of 12 samples from Colorado, 6 of 7 samples from Arizona, and 1 of 5 samples from Texas. PepMV was originally isolated from pepino (Solanum muricatum) in Peru in 1980 (1) and subsequently from tomato in the Netherlands in 1999 (2). To our knowledge, this is the first report of PepMV in North America. References: (1) R. Jones et al. Ann. Appl. Biol. 94:61, 1980. (2) R. A. A. van Vlugt et al. Plant Dis. 84:103, 2000.
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PMID:First Report of Pepino mosaic virus in Canada and the United States. 3082 96