Temperature-dependent actinomycin D effect on RNA synthesis during synchronous development in Neurospora crassa

1971 ◽  
Vol 5 (6) ◽  
pp. 521-532 ◽  
Author(s):  
R. E. Totten ◽  
H. Branch Howe
Keyword(s):  
Neurospora Crassa ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Temperature Dependent ◽  
Synchronous Development

scholarly journals SOME BIOCHEMICAL PROPERTIES OF CHINESE HAMSTER CELLS SENSITIVE AND RESISTANT TO ACTINOMYCIN D

1974 ◽  
Vol 63 (3) ◽  
pp. 773-779 ◽  
Author(s):  
Robert H. F. Peterson ◽  
Judith A. O'Neil ◽  
June L. Biedler
Keyword(s):  
Rna Synthesis ◽  
Actinomycin D ◽  
Chinese Hamster ◽  
Biochemical Properties ◽  
Metabolic Inhibitors ◽  
Drug Resistant ◽  
Temperature Dependent ◽  
Chinese Hamster Cells ◽  
Uptake And Release ◽  
Resistant Cells

A graded series of drug-resistant Chinese hamster sublines has been examined for biochemical changes accompanying resistance to actinomycin D. The most highly resistant subline, DC-3F/AD X, is maintained at 10 µg/ml of the antibiotic. It was shown that over 250 times more actinomycin D is required to inhibit RNA synthesis in this subline than in the parental DC-3F line. The DC-3F/AD X subline was also shown to have a somewhat reduced capacity to transport uridine as compared to parental cells. Sensitive cells took up over 50 times more tritiated antibiotic than the most resistant cells, as determined in a 1-h assay. Uptake of actinomycin D was shown to be temperature-dependent in both resistant and sensitive cells and was not influenced by various metabolic inhibitors. Resistance could not be explained by a rapid uptake and release of the antibiotic, as demonstrated in efflux experiments, or by its metabolism. In addition, highly resistant cells which are cross-resistant to puromycin were shown to have a reduced capacity to take up labeled puromycin. These studies provide further evidence indicating that the mechanism of resistance to actinomycin D is reduced permeability to drug and suggesting that cell membrane alteration accounts for resistance to both actinomycin D and puromycin.

Effect of Some Metabolic Inhibitors on Vinblastine-Induced Ribosomal-Granular Material Complexes

1971 ◽  
Vol 29 ◽  
pp. 548-549
Author(s):  
Awtar Krishan ◽  
Dora Hsu
Keyword(s):  
Granular Material ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Actinomycin D ◽  
Metabolic Inhibitors ◽  
Protein And Rna Synthesis ◽  
Apparent Reduction ◽  
Plant Alkaloids ◽  
In Cells

Cells exposed to antitumor plant alkaloids, vinblastine and vincristine sulfate have large proteinacious crystals and complexes of ribosomes, helical polyribosomes and electron-dense granular material (ribosomal complexes) in their cytoplasm, Binding of H3-colchicine by the in vivo crystals shows that they contain microtubular proteins. Association of ribosomal complexes with the crystals suggests that these structures may be interrelated.In the present study cultured human leukemic lymphoblasts (CCRF-CEM), were incubated with protein and RNA-synthesis inhibitors, p. fluorophenylalanine, puromycin, cycloheximide or actinomycin-D before the addition of crystal-inducing doses of vinblastine to the culture medium. None of these compounds could completely prevent the formation of the ribosomal complexes or the crystals. However, in cells pre-incubated with puromycin, cycloheximide, or actinomycin-D, a reduction in the number and size of the ribosomal complexes was seen. Large helical polyribosomes were absent in the ribosomal complexes of cells treated with puromycin, while in cells exposed to cycloheximide, there was an apparent reduction in the number of ribosomes associated with the ribosomal complexes (Fig. 2).

scholarly journals Biosynthesis of growth hormone in the rat anterior pituitary gland. Stimulation of biosynthesis in vitro by insulin

1973 ◽  
Vol 134 (4) ◽  
pp. 1103-1113 ◽  
Author(s):  
A. Betteridge ◽  
M. Wallis
Keyword(s):  
Growth Hormone ◽  
Anterior Pituitary ◽  
Rna Synthesis ◽  
Glucose Utilization ◽  
Actinomycin D ◽  
Hormone Synthesis ◽  
Pituitary Glands ◽  
Hormone Biosynthesis ◽  
Stimulation Of

The effect of insulin on the incorporation of radioactive leucine into growth hormone was investigated by using rat anterior pituitary glands incubated in vitro. A 50% stimulation over control values was observed at insulin concentrations above 2μm (280munits/ml). The effect was specific for growth hormone biosynthesis, over the range 1–5μm-insulin (140–700munits/ml). Lower more physiological concentrations had no significant effect in this system. Above 10μm (1.4 units/ml) total protein synthesis was also increased. The stimulation of growth hormone synthesis could be partially blocked by the addition of actinomycin D, suggesting that RNA synthesis was involved. Insulin was found to stimulate the rate of glucose utilization in a similar way to growth hormone synthesis. 2-Deoxyglucose and phloridzin, which both prevented insulin from stimulating glucose utilization, also prevented the effect of insulin on growth hormone synthesis. If glucose was replaced by fructose in the medium, the effect of insulin on growth hormone synthesis was decreased. We conclude that the rate of utilization of glucose may be an important step in mediating the effect of insulin on growth hormone synthesis.

Hormone-induced differentiation of antheridial branches in Achlya ambisexualis: dependence on ribonucleic acid synthesis

1974 ◽  
Vol 20 (7) ◽  
pp. 977-980 ◽  
Author(s):  
David K. Horowitz ◽  
Peter J. Russell
Keyword(s):  
Sexual Differentiation ◽  
Ribonucleic Acid ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Steroid Hormone ◽  
Acid Synthesis ◽  
Aquatic Fungus ◽  
Induced Differentiation ◽  
Sexual Steroid ◽  
Achlya Ambisexualis

Sexual differentiation in male strains of the aquatic fungus Achlya ambisexualis Raper is induced by antheridiol, a sexual steroid hormone secreted by female strains. Antheridiol-induced initiation of the morphologically distinct antheridial branches in male Achlya is completely prevented when DNA-dependent RNA synthesis is inhibited by actinomycin D. In addition antheridial branch elongation is inhibited to a degree proportional to the concentration of actinomycin D added. Thus, evidence indicates that RNA synthesis is required for antheridiol-induced initiation of antheridial branching and that continued RNA synthesis is required for elongation of antheridial branches.

Ribosomal RNA synthesis in mitochondria of Neurospora crassa

1973 ◽  
Vol 73 (4) ◽  
pp. 425-437 ◽  
Author(s):  
Yoshiaki Kuriyama ◽  
David J.L. Luck
Keyword(s):  
Neurospora Crassa ◽  
Ribosomal Rna ◽  
Rna Synthesis

scholarly journals Effect of DNA-interacting drugs on phage T7 RNA polymerase.

1998 ◽  
Vol 45 (1) ◽  
pp. 127-132 ◽  
Author(s):  
M Piestrzeniewicz ◽  
K Studzian ◽  
D Wilmańska ◽  
G Płucienniczak ◽  
M Gniazdowski
Keyword(s):  
Rna Polymerase ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
T7 Rna Polymerase ◽  
Distamycin A ◽  
E Coli ◽  
Phage T7 ◽  
Drug Dependent ◽  
Dna Complex ◽  
Kinetics Of

9-Aminoacridine carboxamide derivatives studied here form with DNA intercalative complexes which differ in the kinetics of dissociation. Inhibition of total RNA synthesis catalyzed by phage T7 and Escherichia coli DNA-dependent RNA polymerases correlates with the formation of slowly dissociating acridine-DNA complex of time constant of 0.4-2.3 s. Their effect on RNA synthesis is compared with other ligands which form with DNA stable complexes of different steric properties. T7 RNA polymerase is more sensitive to distamycin A and netropsin than the E. coli enzyme while less sensitive to actinomycin D. Actinomycin induces terminations in the transcript synthesized by T7 RNA polymerase. Despite low dissociation rates of DNA complexes with acridines and pyrrole antibiotics no drug dependent terminations are observed with these ligands.

Synthesis of acid-soluble spore proteins by Bacillus subtilis

1982 ◽  
Vol 152 (3) ◽  
pp. 1117-1125
Author(s):  
J M Leventhal ◽  
G H Chambliss
Keyword(s):  
Bacillus Subtilis ◽  
Maximum Rate ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Sodium Dodecyl ◽  
Molecular Weights ◽  
Spore Proteins ◽  
Major Acid

The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phosphatase synthesis began at about t2, and refractile spores were first observed between t7 and t8. In vivo- and in vitro-synthesized ASSPs comigrated in sodium dodecyl sulfate-polyacrylamide gels. Their molecular weights were 4,600 (alpha and beta) and 11,000 (gamma). The average half-life of the ASSP messages was 11 min when either rifampin (10 micrograms/ml) or actinomycin D (1 microgram/ml) was used to inhibit RNA synthesis.

Einfluß von Actinomycin auf die Vermehrung von Myxoviren

1964 ◽  
Vol 19 (4) ◽  
pp. 316-323 ◽  
Author(s):  
Rudolf Rott ◽  
Christoph Scholtissek
Keyword(s):  
Rna Synthesis ◽  
Actinomycin D ◽  
Viral Rna ◽  
Tissue Cultures ◽  
Virus Rna ◽  
High Concentrations

Actinomycin D (5 γ/ml) inhibits the synthesis of fowl plague virus RNA, S-antigen, hemagglutinin, neuraminidase and infectious particles. Even high concentrations (40 γ/ml) of the antibiotic do not inhibit the synthesis of the corresponding components of NDV. Although purified suspensions of these viruses can be inactivated by actinomycin in the presence of light, this photoeffect does not play an important role in the inhibition of fowl plague virus synthesis. If in the case of fowl plague virus actinomycin is added to tissue cultures at a time after infection when viral RNA synthesis has already started (1½—2 hours p. i.) further synthesis of viral RNA is stopped, while the S-antigen titer continues to rise.

scholarly journals Evidence for the De Novo Synthesis of Erythropoietin in Hypoxic Rats

Blood ◽  
1972 ◽  
Vol 40 (5) ◽  
pp. 662-670 ◽  
Author(s):  
J. C. Schooley ◽  
L. J. Mahlmann
Keyword(s):  
Rna Synthesis ◽  
Actinomycin D ◽  
De Novo ◽  
Protein S ◽  
Male Rats ◽  
Hypoxic Exposure ◽  
De Novo Synthesis ◽  
Serum Erythropoietin

Abstract Significant increases in the serum erythropoietin of male rats occur after the end of a brief hypoxic exposure. These increases in the hormone are almost completely abolished when the kidneys are removed after the hypoxic exposure. Injection of puromycin or cycloheximide after the hypoxic exposure significantly decreases the subsequent increases in serum erythropoietin titers, whereas injections of actinomycin D at this time have no significant effect on erythropoietin levels. Injections of actinomycin D before the hypoxic exposure prevent the increase in serum erythropoietin that normally occurs. These findings suggest that a brief period of hypoxia initiates a DNA-dependent RNA synthesis that regulates the de novo ribosomal synthesis of protein(s) involved in the biogenesis of erythropoietin and that the kidney is essential for these reactions to occur.

scholarly journals Uptake and utilization of mRNA by myogenic cells in culture.

1980 ◽  
Vol 87 (1) ◽  
pp. 65-71 ◽  
Author(s):  
B Mroczkowski ◽  
H P Dym ◽  
E J Siegel ◽  
S M Heywood
Keyword(s):  
Cell Cultures ◽  
Rna Synthesis ◽  
Tissue Specificity ◽  
Kinase Activity ◽  
Actinomycin D ◽  
Creatine Kinase Activity ◽  
Dependent Manner ◽  
In Vitro System ◽  
Myoblast Cell

Primary chick myoblast cultures demonstrate the ability to take up exogenously supplied polyadenylated RNA and express the encoded information in a specific manner. This expression is shown to exhibit tissue specificity. Analysis of creatine kinase activity monitored at various times of incubation in the presence of either polyadenylated or nonpolyadenylated RNA indicates that only the poly(A)+ mRNA is capable of being actively translated. Radioactively labled poly(A)+ mRNA is taken up by the cell cultures in a time-dependent manner and subsequently shown to be associated with polysomes. This association with polysomes does not occur in the presence of puromycin and is unaffected by actinomycin D. Thus, nonspecific interaction with polysomes and induction of new RNA synthesis are ruled out and the association of the exogenously supplied poly(A)+ mRNA with polysomes is indicative of its translation in the recipient cells. When heterologous mRNA (globin) is supplied to the myoblasts, it is also taken up and properly translated. In addition, exogenously supplied myosin heavy chain mRNA is found associated with polysomes consisting of 4-10 ribosomes in myoblast cell cultures while in myotubes it is associated with very large polysomes, thus reflecting the different translational efficiencies that this message exhibits at two very different stages of myogenesis. The results indicate that muscle cell cultures can serve as an in vitro system to study translational controls and their roles in development.

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