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1.
Relationship between uridine kinase activity and rate of incorporation of uridine into acid-soluble pool and into RNA during growth cycle of rat hepatoma cells 总被引:10,自引:0,他引:10
Uridine kinase activity measured in cell-free extracts of Novikoff rat hepatoma cells grown in suspension culture fluctuates about 10 fold during the growth cycle of the cells. Maximum specific activity (units/106 cells) is observed early in the exponential phase and then decreases progressively until the stationary phase. The rate of incorporation of uridine into the acid-soluble pool by intact cells fluctuates in a similar manner and both the rate of uridine incorporation by intact cells and the uridine kinase actvity of the cells increase several fold before cell division commences following dilution of stationary phase cultures with freshmedium. Regardless of the stage of growth, uridine is rapidly phosphorylated to the triphosphate level by the cells. The rates of incorporation of uridine into the nucleotide pool and into RNA by intact cells fluctuate in a similar manner during the growth cycle. However, evidence is presented that indicates that alterations in the rate of incorporation of uridine into RNA are not simply due to changes in the rate of phosphorylation of uridine, but are regulated independently. Inhibition of protein synthesis by treating cells with puromycin or actidione causes a marked inhibition of incorporation of uridine into RNA, but has little effect on the phosphorylation of uridine to UTP for several hours. Thus the depression of incorporation of uridine into RNA probably reflects a decrease in the rate of RNA synthesis as a result of inhibition of protein synthesis. Inhibition of RNA synthesis by treating cells with actinomycin D does not affect the rate of conversion of uridine to UTP and thus results in the accumulation of labeled UTP in treated cells. 相似文献
2.
Inhibition of nucleic acid synthesis in Ehrlich tumor cells by periodate-oxidized adenosine and adenylic acid 总被引:1,自引:0,他引:1
J G Cory M M Mansell C B George D S Wilkinson 《Archives of biochemistry and biophysics》1974,160(2):495-503
Periodate-oxidized adenosine and AMP were inhibitory to both RNA and DNA synthesis in Ehrlich tumor cells in culture. With periodate-oxidized adenosine, the inhibition of RNA synthesis paralleled the inhibition of DNA synthesis. Periodate-oxidized AMP, however, was more inhibitory to DNA synthesis than to RNA synthesis. With both compounds, there was a decrease in the conversion of [14C]cytidine nucleotides to [14C]deoxycytidine nucleotides in the acid-soluble pool. The borohy-dride-reduced trialcohol derivative of the periodate-oxidized adenosine compound was not inhibitory to DNA or RNA synthesis in the tumor cells. The incorporation of [3H]uridine into 28S and 18S ribosomal RNA was inhibited by both periodate-oxidized adenosine and AMP, but the incorporation of [3H]uridine in 45S, 5S, and 4S RNA was essentially unaffected by these compounds. Periodate-oxidized adenosine inhibited Ehrlich tumor cell growth in vivo. 相似文献
3.
4.
Gotteried Galling 《Planta》1971,98(1):50-62
Summary The unicellular green alga Chlorella incorporates labeled uridine mainly into the precursors of chloroplast ribosomes. After treatment with rifampicin for 60 min, the uridine incorporation into the particles is completely inhibited. Chloramphenicol treatment results in the same complete inhibition. In constrast, cycloheximide (actidione) slightly stimulates the incorporation of uridine into the chloroplast ribosome precursors.Short-time incorporation of inorganic phosphate into the ribosome fractions is nearly unaffected by rifampicin and chloramphenicol, but it is strongly inhibited by cycloheximide.Isolation and chromatographic separation of nucleic acids after treatment of cells with rifampicin shows that uridine incorporation into RNA is completely inhibited. Chloramphenicol causes only partial inhibition of uridine labeling in the high molecular weight RNA. Here again, cycloheximide stimulates the uridine incorporation.The results indicate that uridine is preferentially incorporated by Chlorella cells into the chloroplast ribosome precursors. Inorganic phosphate is introduced both into cytoplasmic and into chloroplasmic RNA, but because of the quantitative distribution, the cytoplasmic ribosomes are more extensively labeled. Since only inhibitors of bacterial and chloroplasmic RNA-and protein synthesis affect the formation of uridine-labeled ribosomes, this synthesis must take place in the chloroplast itself.
Abkürzungen DNA Desoxyribonucleinsäure - RNA Ribonucleinsäure - MAK-Säule Säule aus methyliertem Albumin mit Kieselgur - Bis-MSB bis-(O-Methylstyryl)-Benzol - PPO 2,5 Diphenyloxazol - Tris Trimethylaminomethan 相似文献
Abkürzungen DNA Desoxyribonucleinsäure - RNA Ribonucleinsäure - MAK-Säule Säule aus methyliertem Albumin mit Kieselgur - Bis-MSB bis-(O-Methylstyryl)-Benzol - PPO 2,5 Diphenyloxazol - Tris Trimethylaminomethan 相似文献
5.
P G Plagemann 《Journal of cellular physiology》1970,75(3):315-327
At cytostatic concentrations, phenethyl alcohol has immediate and reversible effects on multiple metabolic processes of Novikoff rat hepatoma cells growing in suspension culture. These include an inhibition of the transport of various low molecular weight substances into the cell, an inhibition of DNA and protein synthesis and the processing of ribosomal RNA, and a degradation of ribosomal RNA. All effects might be explained as resulting from an interaction of the chemical with cellular membranes. Phenethyl alcohol does not have an immediate effect on RNA synthesis per se. The immediate failure of phenethyl alcohol-treated cells to incorporate uridine from the medium into RNA is due to an inhibition of the uridine transport reaction. 相似文献
6.
After exposing a line of rat liver epithelial cells to a single dose of the carcinogen N-acetoxy-2-acetylaminofluorene (N-acetoxy-AAF), a dose-dependent decrease in [3H]uridine incorporation into total cellular RNA was found. Approx. 50% inhibition occurred with 0.5 μg/ml of the compound. The kinetics of the response, the effects of actinomycin D, and the fractionation of the newly synthesized RNA by polyacrylamide gel electrophoresis indicated preferential inhibition of the synthesis of 45S ribosomal RNA precursor and a relative sparing of the synthesis of heterogeneous nuclear RNA. 相似文献
7.
Ronald E. Law Ralph M. Sinibaldi Michael R. Cummings Adolph J. Ferro 《Biochemical and biophysical research communications》1976,73(3):600-606
5′-Methylthioadenosine (MTA) inhibits the incorporation of [3H] uridine into RNA in salivary glands of . This effect is not due to an inhibition of [3H] uridine uptake into the glands. The inhibition of RNA synthesis by MTA is concentration dependent and maximum inhibition is observed after 45 minutes of incubation in the presence of 1 mM MTA. Experiments utilizing α-amanitin suggest that the synthesis of heterogeneous RNA is completely inhibited. 相似文献
8.
Inhibition of chloroplast development by tentoxin 总被引:1,自引:0,他引:1
John Bennett 《Phytochemistry》1976,15(2):263-265
Light-dependent chloroplast development in detached pea shoots was measured in terms of chlorophyll synthesis and the synthesis of Fraction 1 protein. Both synthetic processes were inhibited more than 90% by the fungal metabolite, tentoxin (1 or 10 μg/ml). These results place Pisum sativum in the class of tentoxin-sensitive higher plants. Tentoxin, actinomycin D, lincomycin, D-threo-chloramphenicol and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) were compared in their ability to inhibit RNA and protein synthesis by isolated pea chloroplasts. Energy for the synthetic reactions was supplied either by light or by added ATP. Only CCCP gave the same pattern of inhibition as tentoxin, i.e. inhibition of both RNA and protein synthesis in the light-driven system but no inhibition in the ATP-driven system. It is concluded that chloroplast developmental processes are inhibited by tentoxin through the inhibition of photophosphorylation. 相似文献
9.
Lim JS Yoo M Kwon HJ Kim H Kwon YK 《Biochemical and biophysical research communications》2010,394(1):42-47
Currently, [3H]uridine is most often used to monitor rRNA synthesis in cultured cells. We show here that radiolabeled ribonucleoside triphosphates, such as [α-33P]UTP, in culture medium were also incorporated efficiently not only into cells but also into de novo RNA, particularly rRNA. Using this method, we first revealed that endoplasmic reticulum (ER) stress inducers such as tunicamycin and thapsigargin suppressed de novo rRNA synthesis, and that PERK, but not IRE1α or ATF6, mediated the suppression. PERK is known to mediate the suppression of de novo protein synthesis via phosphorylation of eIF2α. Consistently, other translational inhibitors such as PSI, proteasomal inhibitor, and cycloheximide suppressed de novo rRNA synthesis. eIF2α knockdown also suppressed both de novo protein and rRNA syntheses. Furthermore, ER stress reduced cellular ATP levels, and the suppression of rRNA synthesis apparently mitigated their reduction. These observations provided a close link between ATP levels and suppression of de novo rRNA synthesis at ER stress, and we proposed a novel feedback mechanism, in which ATP levels were maintained via suppression of de novo rRNA synthesis in ATP-demanding stresses, such as ER stress. 相似文献
10.
Effect of cordycepin (3''-deoxyadenosine) on virus-specific RNA species synthesized in Newcastle disease virus-infected cells. 总被引:6,自引:6,他引:0
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Cordycepin (3'-deoxyadenosine) has no effect on the size or relative proportions of Newcastle disease virus-specific 18-22S mRNA species nor on the amount or size of the polyadenylic acid associated with them. Cordycepin does, however, cause an inhibition of incorporation of [3H]uridine into 50S virus-specific RNA relative to 18-22S RNA. This inhibition is probably not a direct effect of the drug on the synthesis of 50S viral RNA. Like cycloheximide, another drug which inhibits 50S RNA accumulation in paramyxovirus-infected cells, cordycepin inhibits protein synthesis as measured by amino acid incorporation. It is likely that the inhibition of 50S RNA accumulation is a secondary effect of protein synthesis inhibition. This is supported by the finding that concentrations of cordycepin and cycloheximide, which inhibit protein synthesis to the same extent, have the same effect on the ratio of 50 to 18-22S virus-specific RNA. 相似文献
11.
Cytochalasin B. VI. Competitive inhibition of nucleoside transport by cultured Novikoff rat hepatoma cells 总被引:14,自引:9,他引:5
Cytochalasin B competitively inhibits the transport of uridine and thymidine by Novikoff rat hepatoma cells growing in suspension culture with apparent Ki''s of 2 and 6 µM, respectively, but has no effect on the intracellular phosphorylation of the nucleosides. Choline transport is not affected by cytochalasin B. Results from pulse-chase experiments indicate that cytochalasin B has no direct effect on the synthesis of RNA, DNA, or uridine diphosphate-sugars. The inhibition of uridine and thymidine incorporation into nucleic acids by cytochalasin B is solely the consequence of the inhibition of nucleoside transport. 相似文献
12.
Mechanism of Apparent Transcription Inhibition by Methyl Mercury in Cerebellar Neurons 总被引:1,自引:1,他引:0
Abstract: We have investigated the mechanism of inhibition of RNA synthesis by methyl mercury (MeHg) in isolated neonatal rat cerebellar cells. Each of the three component steps involved in the incorporation of exogenous [3H]uridine into cellular RNA was examined separately in whole-cell and/or subcellular preparations. Nuclear RNA polymerase activity was measured in preparations containing both free nuclei and whole cells. Incorporation of [3H]UTP into nuclear RNA was found to be unimpaired at concentrations of MeHg that inhibited whole-cell incorporation of [3H]uridine by > 75%. Cellular uptake of [3H]uridine was assayed in cerebellar cells treated with KCN to deplete ATP levels and block subsequent phosphorylation reactions of transported uridine. Uptake activity under these conditions was unaffected by MeHg. Measurement of intracellular phosphorylation of [3H]uridine indicated that inhibition of this activity closely paralleled that of RNA synthesis. Quantitation of individual uridine nucleotides by polyethyleneimine-cellulose TLC revealed reduced levels of UTP and UDP whereas levels of UMP were elevated, suggesting that impairment of phosphorylation was not the result of cellular ATP depletion but, more likely, a direct effect on phosphouridine kinase enzymes. This mechanism of MeHg-induced inhibition of RNA synthesis was confirmed by assays of uridine phosphorylation using cell-free extracts in which exogenous ATP was supplied. 相似文献
13.
Noncoordinate control of RNA synthesis in eucaryotic cells 总被引:3,自引:0,他引:3
Inhibition of protein synthesis in confluent monolayers of chick fibroblasts stimulates selectively the synthesis of 4S RNA, resulting in a net accumulation of 4S RNA in the inhibited cells. Under these conditions, inhibition of ribosomal RNA synthesis and processing occurs, as does a decrease in soluble uridine phosphate concentrations; increased pools of certain amino acids are also apparent. Recovery of cells from inhibition is accompanied by a rapidly increasing rate of protein synthesis that lasts for several hours. The small molecular weight RNA synthesized during inhibition of protein synthesis appears properly methylated, and in the presence of cycloheximide and actinomycin D shows a precursor-product conversion. Radiolabeled RNA synthesized during inhibition of protein synthesis is stable following the recovery of cells from inhibition. Stimulation of uridine incorporation into 4S RNA during arrest of protein synthesis is also demonstrated in high-density cultures of L- and Hep-2 cells, suggesting that this non-coordinate stimulation of 4S RNA may be a general property of eucaryotic cells. 相似文献
14.
L Leon M E Vasconcellos W Leon F S Cruz R Docampo W de Souza 《Experimental parasitology》1978,45(2):151-159
Olivacine (a pyridocarbazole derivative) causes ultrastructural and metabolic alterations in Trypanosoma cruzi epimastigotes. The cytoplasm of cells grown in the presence of olivacine appears vacuolated, and swelling of the mitochondria is observed. Concomitant with the ultrastructural alterations, there was a reduction of the respiratory rate as well as inhibition of pyruvate oxidation. A marked inhibition of protein synthesis and a slower although significant inhibition of DNA, RNA, and lipid synthesis were also observed. The in vivo activity of olivacine does not parallel its in vitro effects suggesting inactivation of the drug by the host. 相似文献
15.
Stimulation by abscisic acid of RNA synthesis in discs from healthy and tobacco mosaic virus-infected tobacco leaves 总被引:2,自引:0,他引:2
Uptake of abscisic acid from the culture medium by discs of healthy and tobacco mosaic virus-infected tobacco leaves was measured. Small (two to five-fold) increases in abscisic acid concentration in discs caused increases in rates of [3H]uridine and [3H]adenine incorporation into total nucleic acid, virus RNA and host ribosomal RNA. Net accumulation of virus RNA was also enhanced by abscisic acid. This evidence for stimulation of RNA synthesis is compared with previous reports showing inhibition of RNA synthesis in other tissues. It is suggested that the increase in endogenous abscisic acid caused by tobacco mosaic virus infection may be at least partly responsible for observed increases in rates of RNA synthesis after infection.Abbreviations ABA
abscisic acid
- TMV
tobacco mosaic virus 相似文献
16.
Ross C 《Plant physiology》1974,53(4):635-637
Concentrations of cycloheximide as low as 3 μg/ml inhibited incorporation of labeled orotic acid or uridine into RNA cytidylic acid of soybean (Glycine max) hypocotyl sections. Even lower concentrations of this well known protein synthesis inhibitor interfered with conversion of labeled cytidine into RNA uridylic acid. Both cycloheximide and puromycin inhibited absorption of 3H-phenylalanine and its incorporation into protein, but puromycin did not significantly affect the labeling patterns of RNA cytidylic and uridylic acids when orotic acid-6-14C was fed. Results give further support to the hypothesis that cycloheximide inhibits the interconversion of uridine and cytidine nucleotides, presumably by acting as a glutamine antagonist in the glutamine-dependent reaction catalyzed by cytidine triphosphate synthetase. 相似文献
17.
Uridine transport and RNA synthesis in growing and in density-inhibited animal cells 总被引:19,自引:0,他引:19
Both chick embryo fibroblasts and mouse 3T3 cells reduce the rate at which they incorporate H3 uridine into RNA as their growth becomes inhibited at high cell density. This reduction occurs as a function of the cell population density, and with chick embryo cells (in contrast to 3T3 cells) it is not accompanied by significant medium alterations. This indicates the importance of the cell population density in the control of cellular metabolism. The decline in H3 uridine incorporation is paralleled by a decline in the rate of uptake of the isotope into the acid-soluble pool, suggesting that decreased entry of H3 uridine into the cell, rather than a decreased rate of RNA synthesis, is responsible for the reduced rate of incorporation into RNA of density-inhibited cells. This suggestion was confirmed by finding that when the restriction on uridine uptake was overcome by increasing the concentration of uridine in the medium, the density-dependent inhibition of uridine incorporation was largely reversed. We conclude that, even though the rate of H3 uridine incorporation into RNA is reduced three- to five-fold in density-inhibited cells, the rate of synthesis of pulse-labeled RNA continues at 70 to 85% of the rapidly-growing rate. 相似文献
18.
Benzene may affect hemopoiesis by damaging the bone marrow stroma that provides the microenvironment for hemopoiesis. A possible target of benzene toxicity in the stroma is the macrophage, which is a major source of protein factors required for the proliferation and differentiation of progenitor cells. As an initial approach towards understanding whether benzene inhibits hemopoietic factor production in bone marrow stroma, the metabolism of benzene and phenol has been studied and the effect of benzene and its metabolites on macrophage RNA synthesis has been examined. Benzene is not metabolized in macrophages but phenol, the major metabolite of benzene in bone marrow, is converted by peroxidase in the macrophage to both free metabolites and species which covalently bind to cellular macromolecules. Benzene and its metabolites inhibited RNA synthesis in a dose-dependent manner, with 50% inhibitory concentrations of 5 × 10–3M for benzene, 2.5 × 10–3 M for phenol, 2.5 × 10–5 M for hydroquinone, and 6 × 10–6 M for p-benzoquinone; this inhibition was not attributable to loss of cell viability. Benzene, possibly by an inhibition of uridine transport into macrophages, and phenol, by its conversion to covalently binding species, inhibit RNA synthesis in macrophages and thus may inhibit the synthesis of colony stimulating factors required for hemopoiesis.Abbreviations CFU-G / M
colony forming unit-granulocyte / macrophage
- FCS
fetal calf serum
- IC50
molar concentration causing 50% inhibition
- PBS
phosphate buffered saline 相似文献
19.
RNA extracted from purified rinderpest virus was characterised by sucrose gradient sedimentation and polyacrylamide gel electrophoresis.
The predominant virion RNA species had a sedimentation constant of 46S and its estimated molecular weight was 4.8 × 106 daltons. Consistently high amounts of UMP and AMP were detected. The melting-temperature profile of the virion RNA suggested
absence of secondary structure.
The effect of actionomycin D on the replication of rinderpest virus in Vero cells was studied by following the viral RNA synthesis
using labelled uridine as well as by infectivity titration. The viral RNA synthesis was not affected until 12 h following
infection and was inhibited thereafter between 18 and 48 h to an extent of 25% at 5 and 10 Μg levels of the drug. A 100 to
1000-fold reduction in the infectivity titres was observed in the presence of the drug. These results suggest that actinomycin
D inhibits rinderpest viral RNA replication. Sedimentation analysis of viral RNA extracted from drug-treated cultures showed
inhibition of the genome RNA of rinder-pest virus. 相似文献
20.
B.E. Linebaugh J.A. Rillema 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1982,720(4):346-355
Employing defined media conditions, the insulin sensitivities of mouse mammary gland epithelial cells in primary culture and MCF-7 human mammary epithelial cells were determined. Insulin stimulated the rates of [3H]uridine incorporation into RNA and [3H]leucine incorporation into protein in both primary mouse mammary gland epithelial cell cultures and MCF-7 cell cultures at concentrations approximating the dilution endpoint of the hormone (10−21 M). Insulin stimulated the rate of [3H]thymidine incorporation into DNA in primary mouse mammary gland epithelial cells at the dilution endpoint concentrations. However, MCF-7 cells required insulin concentrations 100–1000-times that necessary in mouse mammary epithelial cultures to elicit an increased rate of [3H]thymidine incorporation into DNA. Evidence is presented which suggests that the increased rates of uptake of [3H]uridine, [3H]thymidine and [3H]leucine into their respective precursor pools is not responsible for the apparent stimulatation of RNA, DNA and protein synthesis. 相似文献