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)

The transcriptional elongation factor TFIIS causes stimulation of RNA polymerase II elongation and readthrough of some of the elongation blocks. We present cloning and sequence characterization of the human TFIIS gene and a pseudogene. The intron-less organization of both of these genes indicates that previously identified cDNAs which suggested the presence of an intron were the products of cloning artifacts. The gene is organized in an uninterrupted ORF which codes for 301 amino acids, whereas the pseudogene lacks an ORF able to code for a full-length protein. The potential promoter for the gene has two putative GC-box-type consensus sequences, two CCAAT-box consensus sequences, and is bounded by a human Alu sequence. Two potential transcriptional termination signal sequences downstream from the consensus polyadenylation signal are proposed.
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PMID:Characterization of the gene encoding the human transcriptional elongation factor TFIIS. 811 16

We demonstrate that the BC200 RNA gene, which encodes a neural small cytoplasmic RNA, is a member of the most prodigious family of interspersed repetitive DNA and that its product represents an example of a primate tissue-specific RNA polymerase III transcript. The BC200 RNA gene is an early monomeric member and one of the few postulated transcriptionally active Alu sequences in this family of nearly half a million retropositionally amplified elements dispersed throughout the human genome. Furthermore, the isolation of two pseudogenes, BC200 beta and BC200 gamma, demonstrates the gene's transpositional ability. Interestingly, the BC200 beta pseudogene may have been generated by a conversion-like event after the human/chimpanzee divergence, resulting in an exchange of the left arm of a dimeric Alu element with the BC200 RNA coding sequence. Our data on conserved features of the active BC200 alpha gene suggest that its RNA product has been "exapted" into a function of the primate brain and provides a selective advantage to the species.
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PMID:BC200 RNA: a neural RNA polymerase III product encoded by a monomeric Alu element. 826 90

In eukaryotes 5S rRNA genes are transcribed by RNA polymerase III. These genes occur in D. discoideum on the ca. 90 copies of an extrachromosomal palindrom together with the other ribosomal RNAs, which are generally transcribed by RNA polymerase I. A 5S rRNA gene has been isolated and its nucleotide sequence as well as the organization of the gene relative to the RNA polymerase I operon has been determined. The sequence of the coding region corresponds exactly to an earlier published 5S rRNA sequence. The genes are located just downstream from the 26S RNA and transcription orientations of the pol I genes and the pol III gene point into the same direction, away from the centromer of the palindrom. The isolated gene appears to be functional since it serves as a specific target for a nuclear protein, most likely TFIIIA. A genomic copy of a 5S rRNA pseudogene has been isolated from the D. discoideum strain V12. This pseudocopy contains nucleotides 52 to 82 of a bona fide 5S rRNA gene with only three mismatches. It resides 78 nucleotides downstream from the glu13(UUC) tRNA gene which in the D. discoideum strain V12 is associated with the retrotransposable element DRE.
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PMID:The Dictyostelium discoideum 5S rDNA is organized in the same transcriptional orientation as the other rDNAs. 846 Oct 13

A reverse genetics approach was applied to generate a chimeric nonsegmented negative strand RNA virus, rabies virus (RV) of the Rhabdoviridae family, that expresses a foreign protein. DNA constructs containing the entire open reading frame of the bacterial chloramphenicol acetyltransferase (CAT) gene and an upstream RV cistron border sequence were inserted either into the nontranslated pseudogene region of a full-length cDNA copy of the RV genome or exchanged with the pseudogene region. After intracellular T7 RNA polymerase-driven expression of full-length antigenome RNA transcripts and RV nucleoprotein, phosphoprotein and polymerase from transfected plasmids, RVs transcribing novel monocistronic mRNAs and expressing CAT at high levels, were recovered. The chimeric viruses possessed the growth characteristics of standard RV and were genetically stable upon serial cell culture passages. CAT activity was still observed in cell cultures infected with viruses passaged for more than 25 times. Based on the unprecedented stability of the chimeric RNA genomes, which is most likely due to the structure of the rhabdoviral ribonucleoprotein complex, we predict the successful future use of recombinant rhabdovirus vectors for displaying foreign antigens or delivering therapeutic genes.
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PMID:Highly stable expression of a foreign gene from rabies virus vectors. 869 89

A VH gene segment that could not be assigned to any of the six known VH gene families of the channel catfish was identified in a genomic clone containing VH gene segments. This gene segment (designated VH7.1) exhibited the structural features characteristic of vertebrate VH genes, specifically potential upstream regulatory sequences, a leader sequence split by an intron, a reading frame that could be readily divided into framework and complementarity determining regions, and a 3' recombination signal sequence. Two regions of nucleotide deletions coupled with degeneracy in the nonamer sequence indicate that this VH gene segment is a pseudogene. Genomic DNA restricted with different enzymes and hybridized under stringent conditions with probes derived from VH7.1 showed that 8-10 bands were present in Southern blots. Reverse transcriptase PCR approaches were used to determine if any of these related sequences were expressed. Sequence analysis of cloned PCR products indicates that different VH gene segments exhibiting > 80% similarity to germline VH7.1 are expressed. Multiple sequence alignments showed that the expressed cVH7a cDNA sequence shared less than 60% nucleotide similarity with representative cDNA sequences from the other known catfish VH gene families. These combined results thus fulfil the criteria for the definition of a new family of catfish VH gene segments. This newly defined, small VH family is designated VH7.
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PMID:Characterization of a seventh family of immunoglobulin heavy chain VH gene segments in the channel catfish, Ictalurus punctatus. 883 18

A two-step reverse-transcriptase-based polymerase chain reaction (PCR) with nested primer pairs was developed to amplify sensitive and specific cytokeratin-19 (CK-19) mRNA sequences from human breast cancer cells. No CK-19 pseudogene interference was seen. The larger DNA-derived amplification products could be clearly discriminated from mRNA-derived products. The CK-19 message was not amplified from bone marrow or blood of healthy volunteers and patients with haematological malignancies nor from myeloid and lymphoid cell lines. Breast cancer cells were diluted in buffy coat cells up to 10(-6) and CK-19 mRNA sought by PCR. The CK-19 message was detected in 14 of 26 blood samples and 14 of 24 marrow samples but in neither of two peripheral blood stem cell samples taken from 35 breast cancer patients. By sequence-analysis control of two of these samples and two cell lines, the amplified DNA fragments were confirmed to be homologous with the CK-19 sequence. The CK-19 message was further sought in matched blood/marrow samples taken from 13 untreated women in the same cohort at the time of diagnosis. In 3 of these, CK-19 RNA was detected in blood and marrow and, in 3 others, only in blood, but never in marrow alone. The results show that CK-19 assay by reverse transcriptase/PCR is a sensitive and specific technique for the detection of cancer cells in bone marrow and blood. It could be helpful in diagnosis and monitoring of metastatic breast cancer and detection of micrometastases. This should be evaluated on larger numbers of patients, with different clinical samples and epithelial malignancies.
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PMID:Reverse transcriptase/polymerase chain reaction detection of cytokeratin-19 mRNA in bone marrow and blood of breast cancer patients. 889 79

A 1.3Mb chromosome 11-specific yeast artificial chromosome (YAC) that spans a t(1;11) translocation breakpoint associated with major psychosis has been used to enrich cDNAs that are encoded within it and expressed in the human foetal brain. Database analysis of the selected fragments led to the identification of 54 clones matching alpha-tubulin, 4 fragments matching two anonymous human expressed sequence tags (ESTs) and 8 fragments giving no database matches. The clones matching alpha-tubulin led to the identification of a novel alpha-tubulin locus located approximately 250 kb proximal to the translocation breakpoint. Extensive sequence and expression analysis of this locus suggests that this is a processed pseudogene, although a long open reading frame is maintained and the possibility that an abnormally acting protein may be expressed in a highly tissue or developmental specific manner cannot be discounted. The novel cDNA fragments map up to 700 kb proximal to the translocation breakpoint and are associated with potential CpG islands. Reverse transcriptase polymerase chain reaction (RT-PCR) expression analysis and high resolution genomic mapping suggest that they may comprise up to three novel genes. No major disruption of the identified fragments could be detected in the genomic DNA of translocation carriers. The psychosis associated with this translocation may therefore be due to position effects on the transcription of these genes or an involvement of translocated chromosome 1 sequences.
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PMID:Novel transcribed sequences neighbouring a translocation breakpoint associated with schizophrenia. 903 12

A human genomic clone designated LhrRAX3 isolated from an X chromosome-specific library was found to have a 28S ribosomal RNA retropseudogene symbolized as RNRP2 within a 12.5-kb human DNA insert. The sequence of the rRNA retropseudogene has an identity of 96% with about 300 nucleotides at the 3'-terminus of the human 28S rRNA gene. RNRP2 is flanked by a pair of perfect direct repeats of 16 nucleotides, the hallmark characteristic of a processed pseudogene having been integrated into the genome. The structural element has a long A-rich tract at its 3'-end, apparently the result of an aberrant polyadenylation event of a RNA polymerase I transcript, prior to its subsequent reverse transcription and retroposition into the genome. An Alu repeat sequence truncated by 80 nucleotides at the 5'-region occurs about 800 base pairs downstream and is of opposite orientation to RNRP2. The Alu element is bounded by 16-nucleotide direct repeats and is a member of the Alu Y subfamily.
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PMID:A human 28S ribosomal RNA retropseudogene. 932 47

Transaldolase (TAL) is a key enzyme of the pentose phosphate pathway, which is responsible for generation of reducing equivalents to protect cellular integrity from reactive oxygen intermediates. While exons 2 and 3 are highly repetitive, the complete TAL-H gene is mapped to a single genomic locus (TALDO1(2)) by several independent approaches. Southern blot hybridization of a 827-bp 3' EcoRI fragment of the TAL-H cDNA to human-mouse somatic cell hybrid DNA localized TALDO1 to the p13-->pter region of chromosome 11. Fluorescence in situ hybridization with a 15-kb genomic fragment harboring exons 1 and 2 mapped TALDO1 to 11p15.4-p15.5. A truncated and mutated segment of TAL-H exon 5 terminating with a poly(A) tail was identified in a pseudogene locus (TALDOP1) on chromosome 1. Reverse transcriptase-PCR studies of human-mouse somatic cell hybrids revealed the presence of the functional TAL-H gene on chromosome 11 and its absence on human chromosome 1. Mapping of radiation hybrids placed TALDO1 between markers WI-1421 and D11S922 on 11p15.
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PMID:The human transaldolase gene (TALDO1) is located on chromosome 11 at p15.4-p15.5. 933 83

In the plastome of the obligate root-parasitic plant, Lathraea clandestina, the rbcL gene has been maintained and is expressed, despite the reduced size and gene content of the plastid genome. Some of the plastid genes involved in translation (e. g. transfer RNAs, ribosomal RNAs and ribosomal proteins) have been sequenced and still appear to code for functional ribosomal components. Indeed, the 16S rRNA and rpl20 genes are expressed whilst other necessary tRNA and ribosomal protein-encoding genes have probably been deleted or truncated. Although obtained by PCR, the four rpo genes for Escherichia coli-like plastid encoded RNA polymerase appear to be pseudogenes. Nevertheless, the rbcL gene, with a "-10, -35" prokaryotic-like promoter, is still transcribed. In contrast to photosynthetic plants, rbcL transcripts in Lathraea are larger in their 5' region and cover the prokaryotic-like promoter. The transcription initiation site is located near the ATG start codon of the atpB pseudogene. Similarity to non-consensus E. coli-like plastid promoters suggests that rbcL transcription is driven by a nuclear-encoded RNA polymerase.
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PMID:The rbcL gene from the non-photosynthetic parasite Lathraea clandestina is not transcribed by a plastid-encoded RNA polymerase. 974 24


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