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)

Mutants of Escherichia coli showing a rifampin-dependent phenotype were isolated from cells of strain CP78 mutagenized with ethyl methane sulfonate or nitrosoguanidine when an antibiotic underlay technique was used. The mutants varied greatly in their rifampin requirement. The minimum necessary concentration ranged from 1 to 50 micrograms/ml. The mutants could be divided into four phenotypic classes. These dependent mutants and their revertants should be a useful tool for probing interactions between the component polypeptides of ribonucleic acid polymerase and for studying the linkage of transcription with other cellular processes.
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PMID:New tool for studying interactions of components of ribonucleic acid polymerase: rifampin-dependent mutants. 38 84

Clones resistant to the cytotoxic action of alpha-amanitin have been isolated from a strain of fetal human lung diploid fibroblasts. Resistant clones were recovered at a frequencey of 5 X 10(-8) after single-step selections following mutagenesis with the mutagen ethyl methane sulfonate. Following propagation in drug-free medium, the clones retained the selected phenotype and in both growth and plating experiments showed a 10-50-fold higher resistance than wild-type cells to the cytotoxicity of 0.25 microgram/ml alpha-amanitin. The alpha-amanitin sensitivity of RNA polymerase II purified from the mutant cells suggests the presence of two forms of the enzyme, one similar to that found in wild-type cells and a second form with increased resistance to alpha-amanitin inhibition. These results are consistent with previous evidence that alpha-amanitin resistance behaves as a codominant marker in mammalian cells.
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PMID:Human diploid fibroblast mutants with altered RNA polymerase II. 102 70

Methylococcus capsulatus (Bath) uses a soluble methane monooxygenase (sMMO) to catalyse the oxidation of methane to methanol. sMMO is comprised of three components; A, B and C. Protein C (the reductase) transfers electrons from NADH to protein A (the hydroxylase) which contains the active site, and protein B regulates this electron flow. The five genes encoding the sMMO proteins and their subunits are clustered and have been cloned in Escherichia coli. A DNA fragment containing mmoB, the gene encoding protein B, was subcloned into pT7-5, a plasmid of the T7 RNA polymerase promoter expression system. Upon induction, E. coli expressed protein B which was fully functional after purification. The gene encoding protein C, mmoC, was amplified with unique restriction sites at each end using the polymerase chain reaction and then subcloned into pT7-7 (a plasmid similar to pT7-5 but containing its own ribosome-binding site and ATG start codon). Protein C expressed in E. coli was also found to be functional. This is the first report of the functional expression of methanotroph methane monooxygenase genes in a heterologous host and represents a significant step forward in our analysis of the assembly and catalysis of sMMO.
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PMID:Functional expression in Escherichia coli of proteins B and C from soluble methane monooxygenase of Methylococcus capsulatus (Bath). 151 60

Methoprene, a chemical analog of juvenile hormone, is toxic when applied to late third-instar larvae of Drosophila melanogaster. Using an ethyl methane sulfonate mutagenesis screen, we have selected two noncomplementing mutants, one of which is nearly 100 times more resistant than wild-type to either methoprene or juvenile hormone III topically applied or incorporated into the diet. The mutation, named methoprene-tolerant (Met), also confers resistance to methoprene-induced pseudotumor formation in larvae as well as to juvenile hormone III- or methoprene-induced vitellogenic oocyte development in adult females. Met adults show little or no cross-resistance to four other insecticides. The mutation was mapped by recombination to a location 35.4 on the X-chromosome and uncovered by chromosomes deficient for the region 10C2-10D4. Complementation was observed between Met and a lethal allele of the RNA polymerase II locus, which is also found in this region. Since the Met mutation also confers resistance to methoprene-induced abnormalities in adult cuticle formation, the autonomy of Met expression could be evaluated in flies mosiac for this mutation. Autonomous expression of Met was found both in abdominal cuticle as well as in external male genitalia. The characteristics of Met are consistent with those expected of a mutant having altered juvenile hormone reception in target tissue.
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PMID:A Drosophila melanogaster mutant resistant to a chemical analog of juvenile hormone. 309 61

Deoxyribonucleic acid-dependent ribonucleic acid polymerase mutants of Bacillus subtilis strain Marburg were isolated after mutagenesis of spores with ethyl methane sulfonate. Genetic analysis by PBS1-mediated transduction and by transformation indicated that mutations responsible for all of the four phenotypic classes studied (rifampin resistance, streptovaricin resistance, streptolydigin resistance, and temperature sensitivity) were clustered close to the cysA14 locus. Three-factor transformation analysis has indicated the most probable marker order as follows: Rif(R)(Stv)(R)-Std(R)-Ts(418)-Ts(427). In addition, further characterization of the classical group I reference marker, cysA14, is reported.
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PMID:Genetic analysis of ribonucleic acid polymerase mutants of Bacillus subtilis. 414 88

To assess the functional role of RNA polymerase II in the regulation of transcription during muscle differentiation, we isolated and characterized a large number of independent alpha-amanitin-resistant (AmaR) mutants of L6 rat myoblasts that express both wild-type and altered RNA polymerase II activities. We also examined their myogenic (Myo) phenotype by determining their ability to develop into mature myotubes, to express elevated levels of muscle creatine kinase, and to synthesize muscle-characteristic proteins as detected by two-dimensional polyacrylamide gel electrophoresis. We found a two- to threefold increase in the frequency of clones with a myogenic-defective phenotype in the AmaR (RNA polymerase II) mutants as compared to control ethyl methane sulfonate-induced, 6-thioguanine-resistant (hypoxanthine, guanine phosphoribosyl transferase) mutants or to unselected survivors also exposed to ethyl methane sulfonate. Subsequent analysis showed that about half of these myogenic-defective AmaR mutants had a conditional Myo(ama) phenotype; when cultured in the presence of amanitin, they exhibited a Myo- phenotype; in its absence they exhibited a Myo+ phenotype. This conditional Myo(ama) phenotype is presumably caused by the inactivation by amanitin of the wild-type amanitin-sensitive RNA polymerase II activity and the subsequent rise in the level of mutant amanitin-resistant RNA polymerase II activity. In these Myo(ama) mutants, the wild-type RNA polymerase II is normally dominant with respect to the Myo+ phenotype, whereas the mutant RNA polymerase II is recessive and results in a Myo- phenotype only when the wild-type enzyme is inactivated. These findings suggest that certain mutations in the amaR structural gene for the amanitin-binding subunit of RNA polymerase II can selectively impair the transcription of genes specific for myogenic differentiation but not those specific for myoblast proliferation.
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PMID:Myogenic differentiation of L6 rat myoblasts: evidence for pleiotropic effects on myogenesis by RNA polymerase II mutations to alpha-amanitin resistance. 686 46

Stable mutants resistant to the nucleoside analog 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB), which interferes with RNA synthesis, have been selected in Chinese hamster ovary (CHO) and human diploid fibroblasts. In CHO cells, upon treatment with the mutagen ethyl-methane sulfonate (EMS), a linear dose--response between the concentration of mutagen and the frequency of DrbR mutants was observed in the range of 20--300 micrograms/ml. The selection system did not show cell density or cross-feeding effects, and the optimal expression time following mutagenesis was found to be 2--3 days for CHO cells and 5--6 days for human fibroblasts. The DrbR mutation behaved codominantly in DrbR x DrbS hybrids. Addition of DRB affected nucleoside uptake to a similar extent in both wild-type and mutant cells, indicating that the drug was able to enter the mutant cells. The failure of DrbR mutants to show any cross-resistance to other toxic nucleoside analogs examined suggests that the action of DRB does not involve the initial phosphorylation step. DRB addition did not cause any marked inhibition of either RNA polymerase I or RNA polymerase II activity from both wild-type and mutant cells in vitro, indicating that its effect on RNA synthesis may be indirect.
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PMID:DRB resistance in Chinese hamster and human cells: genetic and biochemical characteristics of the selection system. 693 Jul 2

Antibiotics can disturb the production of biogas during anaerobic digestion. This study shows a systematic approach to understanding how the different bacterial populations involved in the final conversion of organic matter into methane are inhibited by 15 antimicrobial agents with different specificities and modes of action. The results obtained show the following trends: (i) some inhibitors, such as the macrolide erythromycin, lack any inhibitory effect on biogas production; (ii) some antibiotics, with different specificities, have partial inhibitory effects on anaerobic digestion and decrease methane production by interfering with the activity of propionic-acid- and butyric-acid-degrading bacteria, (e.g. antibiotics that interfere with cell wall synthesis, RNA polymerase activity and protein synthesis, especially the aminoglycosides); (iii) the protein synthesis inhibitors chlortetracycline (IC50 40 mg l-1) and chloramphenicol (IC50 15-20 mg l-1) are very powerful inhibitors of anaerobic digestion. The majority of the antibiotics tested lacked activity against acetoclastic methanogens, being active only on the acetogenic bacteria. However, chloramphenicol and chlortetracycline could cause the complete inhibition of the acetoclastic methanogenic archaea.
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PMID:The action of antibiotics on the anaerobic digestion process. 900 91

RNA polymerase (RNAP) purified from Methanobacterium thermoautotrophicum DeltaH has been shown to initiate transcription accurately in vitro from the hmtB archaeal histone promoter with either native or recombinant forms of the M. thermoautotrophicum TATA-binding protein and transcription factor TFB. Efforts to obtain transcription initiation from hydrogen-regulated methane gene promoters were, however, unsuccessful. Two previously unrecognized archaeal RNAP subunits have been identified, and complex formation by the M. thermoautotrophicum RNAP and TFB has been demonstrated.
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PMID:Methanobacterium thermoautotrophicum RNA polymerase and transcription in vitro. 1040 Jun 4

Genetic experiments have indicated a role for the Ccr4-Not complex in the response to hydroxyurea (HU) induced replication stress and ionizing radiation in yeast. This response includes transcriptional induction of the four genes constituting the ribonucleotide reductase (RNR) enzymatic complex, RNR1-4 and degradation of its inhibitor, Sml1p. The Ccr4-Not complex has originally been described as a negative regulator of RNA polymerase II (pol II) transcription, but it has also been implicated in mRNA turnover and protein ubiquitination. We investigated the mechanism of the HU sensitivity conferred by mutation of CCR4-NOT genes. We found that the ubiquitin protein ligase activity of Not4p does not play a role in HU induced Sml1p degradation. We show, however, that the HU sensitivity of ccr4-not mutant strains correlated very well with a defect in accumulation of RNR2, RNR3 and RNR4 mRNA after HU or methyl-methane sulfonate (MMS) treatment. Chromatin immunoprecipitation (ChIP) experiments show that TBP, pol II and Set1p recruitment to the activated RNR3 locus is defective in cells lacking NOT4. Moreover, RNR3-promoter activity is not induced by HU in these cells. Our experiments show that induction of RNR gene transcription is defective in ccr4-not mutant strains, providing an explanation for their sensitivity to HU.
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PMID:DNA damage and replication stress induced transcription of RNR genes is dependent on the Ccr4-Not complex. 1627 85


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