Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
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Secondary structure of pre-mRNA in nuclear ribonucleoprotein particles (30S-particles) was examined using fluorescent dyes: acridine orange, acriflavine and ethidium bromide. Comparison of ethidium bromide and acriflavine adsorption isotherms for RNP-particles and free RNA and a study of acridine orange dimerization on binding to RNP revealed that 70% of pre-mRNA in 30S-particles is accessible for the dye binding. Dye molecules were adsorbed on double-stranded sequences (11--12% of the total amount of RNA in 30S-particles) and on the single-stranded parts of RNA (58--59% of 30S-particles), the rest part of RNA was unaccessible for the dye binding. A method involving measurements of acriflavine fluorescence quantum yields was used for the determination of nucleotide composition of double-stranded parts of RNA in the 30S-particles. AU-nucleotide content thus obtained was approximately 50%, as was established also for free pre-mRNA. Na+ ions weaken the interaction between the protein and pre-mRNA in 30S particles and increase mobility of double-stranded parts of this nucleic acid.
Mol Biol (Mosk)
PMID:[Nuclear ribonucleoproteins containing messenger RNA. 12. Fluorescence studies of the secondary structure of pre-mRNA in nuclear RNP-particles]. 75 87

The action of ethidium bromide and berenil on the mitochondrial genome of Saccharomyces cerevisiae has been compared in three types of study: (i) early kinetics (up to 4 h) of petite induction by the drugs in the presence or absence of sodium dodecyl sulphate; (ii) genetic consequences of long-term (8 cell generations) exposure to the drugs; (iii) inhibition of mitochondrial DNA replication, both in whole cells and in isolated mitochondria. The results have been interpreted as follows. Firstly, the early events in petite induction differ markedly for the two drugs, as indicated by differences in the short-term kinetics. After some stage a common pathway is apparently followed because the composition of the population of petite cells induced after long-term exposure are very similar for both ethidium bromide and berenil. Secondly, both drugs probably act at the same site to inhibit mitochondrial DNA replication, in view of the fact that a petite strain known to be resistant to ethidium bromide inhibition of mitochondrial DNA replication was found to have simultaneously acquired resistance to berenil. From consideration of the drug concentrations needed to inhibit mitochondrial DNA replication in vivo and in vitro it is suggested that in vivo permeability barriers impede the access of ethidium bromide to the site of inhibition of mitochondrial DNA replication, whilst access of berenil to this site is facilitated. The site at which the drugs act to inhibit mitochondrial DNA replication may be different from the site(s) involved in early petite induction. Binding of the drugs at the latter site(s) is considered to initiate a series of events leading to the fragmentation of yeast mitochondrial DNA and petite induction.
Mol Gen Genet 1975 Dec 09
PMID:Biogenesis of mitochondria. 43. A comparative study of petite induction and inhibition of mitochondrial DNA replication in yeast by ethidium bromide and berenil. 76 29

Yeast tRNAPhe has been modified by excising the Y base and by replacing the Y base or the dihydrouracil residue with the fluorescent dyes proflavine or ethidium bromide. The poly(U) directed ribosomal system from E. coli was used to study the activities of the tRNAPhe derivatives in the assays for ribosome binding and polyphenylalanine synthesis. It was found that the tRNAPhe derivatives modified by replacing dihydrouracil were nearly as active as unmodified tRNAPhe. While excision of the Y base led to a considerable decrease in the activities, replacement by the above mentioned dyes yielded rather active tRNAPhe derivatives. The activities decreased somewhat when the tRNAPhe derivatives with a substitution in the anticodon loop were reduced with NaBH4. The lower plateau values observed with some of the tRNAPhe derivatives are largely attributed to a decreased efficiency of tRNA binding to the ribosomes which, in turn, results in an increased susceptibility to deacylation prior to phenylalanine transfer.
Mol Biol (Mosk)
PMID:[Characterization of fluorescent derivatives of tRNA Phe by experiments in the ribosomal system]. 76 43

When growing cultures of S. cerevisiae are treated with high concentrations of ethidium bromide (greater than 50 mug/ml), three phases of petite induction may be observed: I. the majority of cells are rapidly converted to petite, II. subsequently a large proportion of cells recover the ability to form respiratory competent clones, and III. slow, irreversible conversion of all cells to petite. The extent of recovery of respiratory competence observed is dependent on the strain of S. cerevisiae employed and the temperature and the carbon source used in the growth medium. The effects of 100 mug/ml ethidium bromide are also produced by 10 mug/ml ethidium bromide in the presence of the detergent, sodium dodecyl sulphate, and recovery is also observed when cells are treated with 10 mug/ml ethidium bromide under starvation conditions. Genetic analysis of strain differences indicates that a number of nuclear genes influence petite induction by ethidium bromide. In one strain, S288C, petite induction by 100 mug/ml ethidium bromide is extremely slow under certain conditions. Mitochondria isolated from from S288C lack the ethidium bromide stimulated nuclease activity found in D243-4A, a strain which shows triphasic kinetics of petite formation. This enzyme may, therefore, be responsible for the initial phase of rapid petite formation.
Mol Gen Genet 1976 Mar 30
PMID:Factors affecting petite induction and the recovery of respiratory competence in yeast cells exposed to ethidium bromide. 77 97

The treatment of yeast cells with high levels of ethidium bromide causes a rapid induction of respiratory deficient mutants followed by a period of recovery to respiratory competence in 60 to 70% of the cells. Prolonged exposure then results in a final irreversible phase of petite formation. Sucrose gradient sedimentation analysis of 3H-adenine labelled mtDNA indicates that limited fragmentation (to about 16-18S) occurs during the initial phase of petite induction followed by a reassembly of the fragments during the period corresponding to the recovery of respiratory competence. The reassembly is associated with an ethidium bromide insensitive incorporation of 3H-adenine into mtDNA at a level consistent with repair synthesis. Genetic analyses, based on the transmission of five markers carried on the mtDNA of "repaired rho+" clones, suggests that reassembly occurs with a high degree of fidelity, though in two of a total of twenty five clones differences in marker transmission frequency were observed which could possibly reflect an altered gene order. In addition, a description is given of the marked changes in the suppressive nature of the treated cells and the temporary reduction in the capacity for marker transmission seen to accompany the transitory fragmentation of the mtDNA. The final phase of petite induction is an energy dependent degradation of the mtDNA to produce a rho degrees culture.
Mol Gen Genet 1976 Mar 30
PMID:Molecular and genetic events accompanying petite induction and recovery of respiratory competence induced by ethidium bromide. 77 98

A mutant strain (2-20) isolated by growth on medium containing oligomycin and cycloheximide was also found to be cross resistant to antimyicn, cerulenin, chloramphenicol, tetracycline, triethyltin and triphenylmethylphosphonium bromide, but collaterally sensitive to dequalinium chloride, gentamycin, neomycin, paromomycin and thiolutin. Growth of 2-20, compared to the parental strain and 2 complete revertants, under a variety of environmental conditions revealed that strain 2-20 had an enhanced sensitivity to increased osmolality, elevated pH, and high temperature; in addition, strain 2-20 was unable to polymerize aminoimidazole ribotide at 37 degrees C as shown by the failure to develop a red colony in the presence of ade 2. Four complex solid media (glucose--KCI, galactose, ethanol, ethanol--KCI, Table 1) unable to sustain the growth of strain 2-20 were arbitrarily chosen to monitor cellular growth under different physiological conditions. Tetrad analysis indicated that the complex phenotype (cross resistance, collateral sensitivity, inablity to polymerize aminoimidazole ribotide, absence of growth under adverse physiological conditions) was inherited by an allele of a locus previously shown to result in a permeability barrier of the plasma membrane to chloramphenicol. 582 of 640 subclones used to isolate revertants of 2-20, under four different physiological conditions, were observed to produce a complete revertant of the complex phenotype. It is proposed that the pleiotropic phenotype could result from an alteration of the plasma membrane and mitochondrial inner membrane by a single nuclear gene mutation.
Mol Gen Genet 1976 Mar 30
PMID:Some physiological alteration associated with pleiotropic cross resistance and collateral sensitivity in Saccharomyces cerevisiae. 77 99

It is shown that caffeine antagonizes petite-induction with ethidium bromide under non-growth conditions when administered during or after mutagenic treatment. Caffeine itself is shown to be a petite-inducing agent when cells are grown in liquid glucose-complete-medium in the presence of the drug. A possible mode of action of caffeine in the ethidium bromide induced petite-mutagenesis is discussed.
Mol Gen Genet 1976 Jul 05
PMID:Effect of caffeine on the rho- -induction with ethidium bromide in Saccharomyces cerevisiae. 78 13

Sodium nalidixate inhibited the cell growth and division of several respiratory competent strains of Saccharomyces cerevisiae. A number of cytoplasmic petite strains (both spontaneous and induced by ethidium bromide) were shown to be more resistant to sodium nalidixate than the wild-type strains from which they were derived. There was considerable variation in sensitivity of different petites derived from the same wild-type. Usually petite strains which were induced by ethidium bromide were more resistant than spontaneously arising petites. The susceptibility of a wild-type strain to nalidixate was found to be least when the mitochondrial respiratory system was maximally repressed. It was also noted that sodium nalidixate (100 mug/ml) induced petite mutants.
Mol Gen Genet 1976 Jul 05
PMID:Differential effects of nalidixate on the cell growth of respiratory competent strains and cytoplasmic petite mutants of Saccharomyces cerevisiae. 78 14

1. Retention or loss of mitochondrial markers CR321, OR1, PR454, TR (gene loci RIB1, OLI1, PAR1, TSM1 respectively has been analysed in a large number of ethidium bromide induced primary rho-clones. Retention of one or more of the four markers with a single clone was observed frequently, only 20 to 25% of clones were found to be (TOCOOOPO). Primary clones retaining two or more of the four markers were found to be mixed, i.e. the primary rho- cell contained a heterogeneous population of variously deleted mitDNA molecules which segregated into different cell lines in the corresponding primary clone. 2. A representative sample of the population of ethidium bromide induced rho- mutants has been analysed by a first subcloning performed after some 30 cell generations of vegetative multiplication in the abscence of the drug. At this level the heterogeneous population of mitDNA molecules, generated by the mutagenic treatment in the primary cell, has been sorted out. The cells forming secondary clones are thus essentially homoplasmic. In contrast to primary clones, genotypes of secondary clones therefore could be determined unambiguously, and the frequency of cell types can be regarded as a faithful representation of the frequency of mitDNA molecules. Retention of markers was low, in less than 2% of secondary clones one or several markers have been found. This observation has been interpreted as indicating that induction of rho-mutants by ethidium bromide is accompanied by deletion of very large sequences of mitDNA in a very large fraction of mitDNA molecules. 3. Five individual rho-clones retaining the four markers TRCRORPR have been isolated and analysed for spontaneous deletion of one or several of these markers during successive subclonings (pedigree analysis). High genetic stability (98-99.5% per cell generation) has been observed in these clones. 4. A method has been developed allowing an unambiguous determination of the order of the four markers on a circular map. It is based on the concomitant loss of two markers and retention of the other two markers (double loss/double retention analysis). The results of four out of five pedigrees of individual rho-clones analysed (spontaneous deletion) and the results of the analysis of populations of secondary rho-clones (ethidium bromide induced deletion) were in full agreement and the order of genes has been determined as being P-T-C-O-P. In the fifth pedigree results suggest an inversion of the T and C markers. 5. Relative distances between pairs of markers have been derived from the frequencies of separation of markers by deletion and were found to be C-T less than C-O less than T-O less than T-P less than C-P less than O-P. Linkage of the four markers could be established, and distances calculated are additive. 6. The general relevance of this approach of mapping by deletion and the methods used for the determination of order and distances of mitochondrial genes has been discussed. (ABSTRACT TRUNCATED)
Mol Gen Genet 1976 Jul 23
PMID:Mapping of mitochondrial genes in Saccharomyces cerevisiae. Populations and pedigree analysis of retention or loss of four genetic markers in Rho-cells. 78 15

Three antimycin resistant mutants of Saccharomyces cerevisiae are characterized genetically. The mutations have been shown to be cytoplasmically inherited by four criteria. The phenotype persists in diploids formed by a cross with a pO strain of yeast of the opposite mating type. Diploids heterozygous for the antimycin marker, however, show segregation of the resistance and sensitivity during mitosis. Tetrad analysis indicates a non-Mendelian segregation (4:0 and 0:4) of the mutations. The antimycin marker can be eliminated by ethidium bromide treatment under conditions that should have deleted all of the mitochondrial DNA.
Mol Gen Genet 1976 Jul 23
PMID:Cytoplasmic inheritance of antimycin A resistance in Saccharomyces cerevisiae. 78 16


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