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)

Anti-HIV drugs act by blocking intracellular replication of the virus by inhibiting viral enzyme either reverse transcriptase or protease. Reverse transcriptase inhibitors belong to 3 different categories: nucleoside analogues, which active forms are the triphosphorylated intracellular compound, non nucleoside inhibitors and the more recent nucleotide analogues. Protease inhibitors are very active in vitro but besides their digestive side effects, have numerous drug interactions because they are metabolised through P450 liver cytochromes, and are associated with long-term toxicity.
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PMID:[Anti-HIV drugs]. 1057 7

Reverse transcriptase-polymerase chain reaction was used to amplify a partial cDNA from rabbit lung mRNA that shared 77% protein sequence identity with the mouse pregnane X receptor (PXR). Rapid amplification of cDNA ends from a rabbit kidney lambdaZAP expression library resulted in the isolation of overlapping cDNAs spanning the complete coding sequence. The deduced amino acid sequence of 411 residues exhibited 79% overall amino acid identity with human PXR and 77% identity with mouse PXR. Based on this protein sequence relationship and a similar degree of conservation exhibited by the mouse and human PXR orthologs, the cDNA appears to encode the rabbit PXR ortholog. 5'-rapid amplification of cDNA ends performed on an adaptor-ligated cDNA library from rabbit liver revealed the presence of an alternate mRNA, which differed at the 5'-terminus. RNase protection assays indicated that the alternate mRNA was expressed at >50-fold lower levels in rabbit kidney and liver. Rifampicin treatment of CV-1 cells cotransfected with a rabbit PXR expression plasmid and a luciferase reporter construct containing two copies of the DR3 enhancer from CYP3A23 produced a 6-fold induction of luciferase activity. In contrast, rat PXR was not responsive to this antibiotic under the same conditions. Pregnenolone 16alpha-carbonitrile was an efficacious activator of rat PXR, but failed to significantly activate rabbit PXR at equivalent concentrations. These results indicate that the ligand activation profile of rabbit PXR is distinct from rat PXR and more closely resembles that of human PXR. The rabbit PXR activation profile is consistent with the cytochrome P450 (P450) 3A6 induction profile in rabbits.
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PMID:Rabbit pregnane X receptor is activated by rifampicin. 1077 31

A cDNA encoding a new cytochrome P450 was isolated from a mouse brain library. Sequence analysis reveals that this 1,958-base pair cDNA encodes a 57-58-kDa 502-amino acid polypeptide that is 70-91% identical to CYP2J subfamily P450s and is designated CYP2J9. Recombinant CYP2J9 was co-expressed with NADPH-cytochrome P450 oxidoreductase (CYPOR) in Sf9 cells using a baculovirus system. Microsomes of CYP2J9/CYPOR-transfected cells metabolize arachidonic acid to 19-hydroxyeicosatetraenoic acid (HETE) thus CYP2J9 is enzymologically distinct from other P450s. Northern analysis reveals that CYP2J9 transcripts are present at high levels in mouse brain. Mouse brain microsomes biosynthesize 19-HETE. RNA polymerase chain reaction analysis demonstrates that CYP2J9 mRNAs are widely distributed in brain and most abundant in the cerebellum. Immunoblotting using an antibody raised against human CYP2J2 that cross-reacts with CYP2J9 detects a 56-kDa protein band that is expressed in cerebellum and other brain segments and is regulated during postnatal development. In situ hybridization of mouse brain sections with a CYP2J9-specific riboprobe and immunohistochemical staining with the anti-human CYP2J2 IgG reveals abundant CYP2J9 mRNA and protein in cerebellar Purkinje cells. Importantly, 19-HETE inhibits the activity of recombinant P/Q-type Ca(2+) channels that are known to be expressed preferentially in cerebellar Purkinje cells and are involved in triggering neurotransmitter release. Based on these data, we conclude that CYP2J9 is a developmentally regulated P450 that is abundant in brain, localized to cerebellar Purkinje cells, and active in the biosynthesis of 19-HETE, an eicosanoid that inhibits activity of P/Q-type Ca(2+) channels. We postulate that CYP2J9 arachidonic acid products play important functional roles in the brain.
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PMID:Cytochrome P450 CYP2J9, a new mouse arachidonic acid omega-1 hydroxylase predominantly expressed in brain. 1132 10

Arabinose has been serendipitously observed to enhance the expression of P450s in Escherichia coli. To understand the mechanism of this arabinose-dependent enhancement, the effects of various carbohydrates were investigated. Surprisingly, a series of sugars, including pentoses and hexoses, enhanced the foreign gene expression in a manner similar to arabinose. Furthermore, glycerol, a poor carbon source, also enhanced P450 expression. These results indicate that the enhancement is independent of the specific efficiency of the carbon source and also suggest the involvement of osmotic stress. Therefore, the effect of the sigma(s) (also termed sigma(38)) factor, a sigma subunit of RNA polymerase that plays a central role in regulating the expression of osmotic stress response genes, has been examined. We found that the glycerol-dependent increase in P450 expression was not observed in sigma(s)-deficient E. coli, indicating that carbohydrates enhance the foreign gene expression in E. coli via the induction of the osmotic stress response. The results suggest the important role of the osmotic stress response in posttranscriptional processes required for producing functional proteins.
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PMID:Osmotic stress induced by carbohydrates enhances expression of foreign proteins in Escherichia coli. 1155 16

Many carcinogens exert their cancer-causing effects by reacting with DNA either directly or following metabolic activation, resulting in covalently linked combination molecules known as carcinogen-DNA adducts. The presence of such lesions in the genome increases the error frequency of the replication machinery, causing mutations that contribute to the initiation and progression of cancer. Cellular DNA repair pathways remove carcinogen adducts from DNA, thus averting the mutagenic potential of many DNA lesions by reducing their presence in the genome. Bulky DNA adducts, like those derived from a number of activated environmental carcinogens such as polycyclic aromatic hydrocarbons (PAHs), are primarily repaired by the nucleotide excision repair (NER) pathway. Transcription-coupled NER (TC-NER) preferentially removes lesions from the transcribed strand of actively expressed genes, and RNA polymerase II stalled at the lesion quite possibly initiates the pathway. Among the bulky DNA adducts that are subject to TC-NER are those resulting from the reaction of the metabolically activated PAH benzo[a]pyrene (BP) with DNA. The P450 mixed-function oxygenases convert BP into a number of reactive intermediates, including tumorigenic (+)- and non-tumorigenic (-)-anti-benzo[a]pyrene diol epoxide (BPDE) that react with DNA via trans epoxide opening to form (+)-trans-anti-[BP]-N(2)-dG ((+)-ta[BP]G) and (-)-trans-anti-[BP]-N(2)-dG ((-)-ta[BP]G), respectively. To test the effect of these lesions on RNA synthesis, in vitro transcription assays using human nuclear extracts were performed with DNA templates containing an RNAPII promoter and a stereochemically pure (+)- or (-)-ta[BP]G adduct on the transcribed or non-transcribed strand. Transcription past (+)- or (-)-ta[BP]G adducts was investigated in the same sequence context to examine stereochemical effects. The (+)-ta[BP]G adduct was investigated in two different local sequence contexts to determine if the surrounding bases influence the adduct's ability to block transcription. These experiments revealed that (+)- and (-)-ta[BP]G adducts on the transcribed strand of the DNA template block RNAPII in a sequence and stereochemistry-dependent manner; however, adducts on the non-transcribed strand do not block elongation significantly but may increase pausing at innate pause sites. In order to elucidate biologically influential differences between the (+)- and (-)-ta[BP]G structures, the DUPLEX program was used to carry out potential energy minimization searches at model transcription junctions. The lowest-energy minimum for the (+)-ta[BP]G adduct gives a structure in which the benzo[a]pyrenyl ring system resides in the minor groove of the heteroduplex region. In contrast, the lowest-energy minimum for a (-)-ta[BP]G adduct shows an orientation in which the benzo[a]pyrenyl group adopts a carcinogen/base-stacked conformation. These conformational preferences may contribute to the differential treatment of (+)- and (-)-ta[BP]G adducts by human RNAPII. In addition, while previous experiments showed that BPDE adducts cause T7RNAP to produce a ladder of truncated transcripts, RNAPII is blocked entirely at only one or two positions by the (+)- and (-)-ta[BP]G adducts, depending on sequence context. It is likely that these differences between the behaviors of T7RNAP and human RNAPII are a result of the structural characteristics of the enzymes' active sites, a hypothesis that is explored in light of their known crystal structures.
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PMID:DNA adducts from a tumorigenic metabolite of benzo[a]pyrene block human RNA polymerase II elongation in a sequence- and stereochemistry-dependent manner. 1213 31

Changes in gene expression regulated by peroxisome proliferator-activated receptor gamma (PPARgamma) and in gene expression related to the inhibin/activin-follistatin system in the rat testis induced by a single oral administration of di-n-butyl phthalate (DBP) (8.6 mmol/kg) were examined and compared with those in the control rats using reverse-transcriptase polymerase chain reaction (RT-PCR). The increase in cytochrome P450 4A1 mRNA, which is regulated by PPARalpha, was significant, but not so profound as the increase of P450 4A1 mRNA in the liver. In contrast, a remarkable increase in the mRNA level of plasminogen activator inhibitor-1 (PAI-1) was found in the testis, suggesting the activation of PPARgamma. The substantial increase in PAI-1 may be related to the disruption of spermatogenesis. On the other hand, significant suppression of the mRNA level of inhibin beta(B) and elevation in the mRNA level of follistatin, an activin-binding protein, were observed after the DBP-administration. Activin B, a homodimer of inhibin beta(B), is known to stimulate spermatogonial proliferation. The present results suggest that the suppression of spermatogenesis resulting from the changes in the expression of genes involved in the inhibin/activin-follistatin system is one of the mechanisms of the testicular atrophy induced by DBP.
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PMID:Changes in peroxisome proliferator-activated receptor gamma-regulated gene expression and inhibin/activin-follistatin system gene expression in rat testis after an administration of di-n-butyl phthalate. 1256 98

The role of the putative P450 monooxygenase OxyD and the chlorination time point in the biosynthesis of the glycopeptide antibiotic balhimycin produced by Amycolatopsis balhimycina were analyzed. The oxyD gene is located directly downstream of the bhp (perhydrolase) and bpsD (nonribosomal peptide synthetase D) genes, which are involved in the synthesis of the balhimycin building block beta-hydroxytyrosine (beta-HT). Reverse transcriptase experiments revealed that bhp, bpsD, and oxyD form an operon. oxyD was inactivated by an in-frame deletion, and the resulting mutant was unable to produce an active compound. Balhimycin production could be restored (i) by complementation with an oxyD gene, (ii) in cross-feeding studies using A. balhimycina JR1 (a null mutant with a block in the biosynthesis pathway of the building blocks hydroxy- and dihydroxyphenylglycine) as an excretor of the missing precursor, and (iii) by supplementation of beta-HT in the growth medium. These data demonstrated an essential role of OxyD in the formation pathway of this amino acid. Liquid chromatography-electrospray ionization-mass spectrometry analysis indicated the biosynthesis of completely chlorinated balhimycin by the oxyD mutant when culture filtrates were supplemented with nonchlorinated beta-HT. In contrast, supplementation with 3-chloro-beta-HT did not restore balhimycin production. These results indicated that the chlorination time point was later than the stage of free beta-HT, most likely during heptapeptide synthesis.
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PMID:Biosynthesis of chloro-beta-hydroxytyrosine, a nonproteinogenic amino acid of the peptidic backbone of glycopeptide antibiotics. 1534 78

Fusarium head blight (FHB), caused by species of the fungus Fusarium, is a worldwide disease of wheat (Triticum aestivum L.). The Chinese T. aestivum 'Ning7840' is one of few wheat cultivars with resistance to FHB. To identify differentially expressed genes corresponding to FHB resistance, a cDNA library was constructed using pooled mRNA isolated from glumes of 'Ning7840' harvested at 2, 6, 12, 24, 36, 72, and 96 h after inoculation (hai) with a conidia spore suspension of Fusarium graminearum. Suppressive subtractive hybridization (SSH) cDNA subtraction was carried out using pooled glume mRNAs from the tester and the control. The cDNA library was differentially screened using the forward subtracted cDNAs and the reverse subtracted cDNAs as probes. Twenty-four clones with significant matches to either plant (16 sequences) or fungal (8 sequences) genes were isolated based on their specific hybridization with forward subtracted cDNA and not reverse subtracted cDNA. Six putative defense-related genes were confirmed by real-time quantitative reverse-transcriptase PCR. Many-fold higher induction of three clones (A3F8, B10H1, and B11H3) in the resistant genotypes compared with susceptible genotypes indicates a putative role in the resistance response to Fusarium graminearum. Transcript accumulations of P450, chitinase (Chi1), and one unknown gene (clone B8Q9) in both resistant and susceptible genotypes suggest an involvement in a generalized resistance response to F. graminearum. Nucleotide sequence analysis showed that cDNA clone A4C6 encodes a cytochrome P450 gene (CYP709C3v2), including 14 N-terminal amino acids that have a membrane-associated helical motif. Other domains characteristic of eukaryotic P450 are also present in CYP709C3v2. The deduced polypeptide of cDNA clone B2H2 encodes an acidic isoform of class I chitinase containing a 960-bp coding region. Southern hybridization using aneuploid lines of T. aestivum 'Chinese Spring' indicated that CYP709C3v2 was located on the short arm of chromosomes 2B and 2D.
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PMID:Induction of wheat defense and stress-related genes in response to Fusarium graminearum. 1572 94

Steroid P450 11beta-hydroxylase, encoded by the CYP11B gene, is a key mitochondrial enzyme for the production of 11-oxygenated androgens, which have been shown to be potent masculinising steroids in several fish species. In this study we have isolated a CYP11B cDNA of 1903 base pairs from the testis of European sea bass (Dicentrarchus labrax) encoding a predicted protein of 552 amino acids. The amino acid identities to other vertebrate 11beta-hydroxylase proteins ranged from 66% to 72% to other fish; 45% to amphibian and 35-39% to mammalian. Southern blot indicated that a single CYP11B gene is present. Northern blot analysis detected two transcripts in testis and head kidney, one of the same size of the isolated clone and the other of 3.9 kb. Reverse transcriptase-polymerase chain reaction showed abundant mRNA expression only in testis and head kidney, being residual in a range of other tissues. Expression of CYP11B and CYP19A (which encodes for ovarian aromatase) was detected from at least 4 days post-hatching and did not appear to be affected by rearing temperature (15 and 20 degrees C) during the first 60 days, a period in which high temperatures promote masculinisation in European sea bass. Throughout, gonadogenesis (60-300 dph), a highly dimorphic pattern of CYP11B expression was consistent with a role of this gene in testicular development.
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PMID:A cDNA for European sea bass (Dicentrachus labrax) 11beta-hydroxylase: gene expression during the thermosensitive period and gonadogenesis. 1696 21

Rifalazil is a benzoxazinorifamycin which inhibits bacterial DNA-dependent RNA polymerase. The benzoxazine ring endows benzoxazinorifamycins with unique physical and chemical characteristics which favor the use of rifalazil and derivatives in treating diseases caused by the obligate intracellular pathogens of the genus chlamydia. Minimal inhibitory concentrations of benzoxazinorifamycins against chlamydia are in the pg/mL range. These compounds have potential as monotherapeutic agents to treat chlamydia-associated disease because they retain activity against chlamydia strains resistant to currently approved rifamycins such as rifampin. A pivotal clinical trial with rifalazil has been initiated for the treatment of peripheral arterial disease. The rationale for this innovative use of rifalazil, including the association of C. pneumoniae in atherosclerotic plaque formation, as well as rifalazil's potency and efficacy against chlamydia in both preclinical and clinical studies, is discussed. Other benzoxazino derivatives may have utility as stand-alone topical antibacterials or combination antibacterials to treat serious Gram-positive infections. None of the benzoxazinorifamycins examined to date induce the cytochrome P450 3A4 enzyme. This is in contrast to currently approved rifamycins which are strong inducers of P450 enzymes, resulting in drug-drug interactions that limit the clinical utility of this drug class.
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PMID:Rifalazil and other benzoxazinorifamycins in the treatment of chlamydia-based persistent infections. 1791 77


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