scholarly journals Deoxyribonucleic acid-dependent ribonucleic acid polymerases from normal and polyoma-transformed BHK-21/C13 cells

1975 ◽  
Vol 145 (3) ◽  
pp. 509-516 ◽  
Author(s):  
R J Cooper ◽  
H M Keir
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Dna Content ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Activity Ratio ◽  
Cell Types ◽  
Dna Templates ◽  
Alpha Amanitin

DNA-dependent RNA polymerase (EC 2.7.7.6) ACTIVITIES FROM NORMAL BHK-21/C13 cells and from BHK-21/C13 cells transformed by polyoma virus (PYY cells) were solubilized and fractionated on columns of DEAE-Sephadex. Various properties of the A and B enzymes from the two types of cell were compared. 1. The yields of polymerase relative to the DNA content of the nuclear preparations are similar for both cell types. 2. The ionic-strength optima of polymerases A and B are 12.5 mM and 100mM with respect to (NH4)2SO4 for both cell types. 3. The Mn2+/Mg2+ activity ratio (measured at the respective optimum for each cation) for polymerase A from BHK-21/C13 cells was 1.48 and for the polymerase A from PYY cells was 0.55. The corresponding ratios for polymerase B were 10.11 for BHK-21/C13 cells and 22.75 for PYY cells. 4. Minor differences in the ability of the A polymerases to transcribe native and denatured DNA templates were observed; such differences were not apparent when the B polymerases were compared. 5. All the polymerases were inhibited completely by actinomycin D and by rifampicin AF/013, but not markedly so by rifampicin. Alpha-amanitin inhibited polymerase B but not polymerase A.

scholarly journals Effect of an exotoxin from Bacillus thuringiensis on Deoxyribonucleic acid-dependent ribonucleic acid polymerase in nuclei from adult Sarcophaga bullata. Unusual behaviour of eukaryotic polymerases to inhibitors

1973 ◽  
Vol 136 (1) ◽  
pp. 9-13 ◽  
Author(s):  
T. J. C. Beebee ◽  
R. P. M. Bond
Keyword(s):  
Ionic Strength ◽  
Bacillus Thuringiensis ◽  
Rna Polymerase ◽  
Metal Ions ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Low Concentration ◽  
Sarcophaga Bullata ◽  
Unusual Behaviour

The DNA-dependent RNA polymerase activities in nuclei isolated from adult Sarcophaga bullata are unusual in their responses to metal ions, ionic strength and inhibitors. There is an activity that is sensitive both to rifamycin and to α-amanitin. The activity is less sensitive to Bacillus thuringiensis exotoxin than is larval polymerase, and low concentration of exotoxin provoke a slight stimulation.

scholarly journals A procedure for the rapid purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase (Short Communication)

1973 ◽  
Vol 133 (1) ◽  
pp. 201-203 ◽  
Author(s):  
Peter Humphries ◽  
David J. McConnell ◽  
Robert L. Gordon
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Short Communication ◽  
Gel Filtration ◽  
High Salt ◽  
Rapid Procedure ◽  
Complete Purification ◽  
Rapid Purification

A rapid procedure involving DNA–cellulose chromatography followed either by sedimentation in a high-salt glycerol gradient or by gel filtration is described for the complete purification of Escherichia coli DNA-dependent RNA polymerase.

scholarly journals Effects of α-amanitin on the stimulation of prostatic ribonucleic acid polymerase by prostatic steroid–protein receptor complexes

1974 ◽  
Vol 140 (3) ◽  
pp. 565-567 ◽  
Author(s):  
P. Davies ◽  
K. Griffiths
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Ribonucleic Acid ◽  
Selective Inhibitor ◽  
High Ionic Strength ◽  
Receptor Complexes ◽  
Protein Receptor ◽  
Stimulation Of

Stimulation of prostatic RNA polymerase in vitro by prostatic 17β-hydroxy-5α-androstan-3-one (5α-dihydrotestosterone)–receptor complexes has been previously reported. By use of the selective inhibitor, α-amanitin, we have shown that both nucleolar and extranucleolar RNA polymerase activities may be stimulated, but stimulation is abolished at high ionic strength.

scholarly journals In vitro initiation of transcription by RNA polymerase II on in vivo-assembled chromatin templates.

1992 ◽  
Vol 12 (4) ◽  
pp. 1639-1651 ◽  
Author(s):  
S C Batson ◽  
R Sundseth ◽  
C V Heath ◽  
M Samuels ◽  
U Hansen
Keyword(s):  
Transcription Factors ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Viral Dna ◽  
Dna Templates ◽  
Initiation Of Transcription ◽  
Transcription In Vitro ◽  
Alpha Amanitin

We have studied the initiation of transcription in vitro by RNA polymerase II on simian virus 40 (SV40) minichromosomal templates isolated from infected cells. The efficiency and pattern of transcription from the chromatin templates were compared with those from viral DNA templates by using two in vitro transcription systems, either HeLa whole-cell extract or basal transcription factors, RNA polymerase II, and one of two SV40 promoter-binding transcription factors, LSF and Sp1. Dramatic increases in numbers of transcripts upon addition of transcription extract and different patterns of usage of the multiple SV40 initiation sites upon addition of Sp1 versus LSF strongly suggested that transcripts were being initiated from the minichromosomal templates in vitro. That the majority of transcripts from the minichromosomes were due to initiation de novo was demonstrated by the efficient transcription observed in the presence of alpha-amanitin, which inhibited minichromosome-associated RNA polymerase II, and an alpha-amanitin-resistant RNA polymerase II, which initiated transcription in vitro. The pattern of transcription from the SV40 late and early promoters on the minichromosomal templates was similar to the in vivo pattern of transcription during the late stages of viral infection and was distinct from the pattern of transcription generated from viral DNA in vitro. In particular, the late promoter of the minichromosomal templates was transcribed with high efficiency, similar to viral DNA templates, while the early-early promoter of the minichromosomal templates was inhibited 10- to 15-fold. Finally, the number of minichromosomes competent to initiate transcription in vitro exceeded the amount actively being transcribed in vivo.

scholarly journals Decreased ribonucleic acid synthesis in isolated rat liver nuclei during starvation

1970 ◽  
Vol 120 (2) ◽  
pp. 381-384 ◽  
Author(s):  
D. Rickwood ◽  
H. G. Klemperer
Keyword(s):  
Rna Polymerase ◽  
Ammonium Sulphate ◽  
Ribonucleic Acid ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Acid Synthesis ◽  
Isolated Nuclei ◽  
Isolated Rat Liver ◽  
Liver Nuclei ◽  
Isolated Rat

1. Isolated nuclei from starved rats showed a lowered incorporation of [14C]UMP into RNA. 2. The Mg2+-dependent incorporation was decreased by 30% after 1 day of starvation, but incorporation in the presence of Mn2+ and ammonium sulphate decreased only after longer periods of starvation. 3. RNA synthesis by nuclei in the presence of excess of added RNA polymerase was unchanged after 1 day of starvation and was inhibited by 20% after 4 days. 4. The capacity of nuclei to bind actinomycin D was unchanged after 1 day and was decreased by 20% after 4 days of starvation.

More evidence for replication-transcription-coupling in Physarum polycephalum

1980 ◽  
Vol 41 (1) ◽  
pp. 105-113
Author(s):  
G. Pierron ◽  
H.W. Sauer
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Physarum Polycephalum ◽  
Mitotic Cycle ◽  
Polymerase Activity ◽  
S Phase ◽  
Isolated Nuclei ◽  
Rna Polymerase Activity ◽  
High Level ◽  
Alpha Amanitin

Endogenous RNA polymerase activity of isolated nuclei from Physarum polycephalum was determined at high (400 mM KCl) and low (5–100 mM KCl) ionic strength. The activity of RNA polymerase B (alpha-amanitin-sensitive UMP incorporation) and of RNA polymerase A (plus C) (alpha-amanitin-resistant UMP incorporation) was compared in accurately sized nuclear samples derived from macroplasmodia at distinct points of the mitotic cycle. Minimum total RNA polymerase activity was detected in metaphase nuclei. A constant level of RNA polymerase B activity was detected at all other stages of the mitotic cycle, if nuclei were assayed at high ionic strength. However, a high level in S-phase, a low level in G2-phase and again a high level in early prophase were measured, if nuclei were assayed at low ionic strength. Inhibition of DNA synthesis by hydroxyurea in vivo had a selective and drastic effect on in vitro RNA polymerase activity of isolated nuclei derived from S-phase plasmodia, yielding up to 100% inhibition in early S-phase.

scholarly journals Use of actinomycin D for the specific quenching of fluorescence of deoxyribonucleic acid in cells stained with acridine aminoderivatives.

1976 ◽  
Vol 24 (11) ◽  
pp. 1169-1172 ◽  
Author(s):  
A V Zelenin ◽  
E A Kirianova ◽  
V A Kolesnikov ◽  
N G Stepanova
Keyword(s):  
Acridine Orange ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Quenching Of Fluorescence ◽  
Approximate Estimation ◽  
In Cells

Actinomycin D specifically quenches the fluorescence of acridine orange and quinacrine bound to deoxyribonucleic acid in cytologic preparations, but does not change the fluorescence of these fluorochromes bound to RNA. The following fluorescence-cytochemical applications of techniques based on these findings can be suggested: (a) distinction between deoxyribonucleic acid and ribonucleic acid; (b) detection of double-stranded virus ribonucleic acid; (c) approximate estimation of the lengths of A-T sequences in deoxyribonucleic acid molecules.

The mechanism of action of griseofulvin

1968 ◽  
Vol 14 (2) ◽  
pp. 111-118 ◽  
Author(s):  
Floyd M. Huber ◽  
David Gottlieb
Keyword(s):  
Cell Wall ◽  
Botrytis Cinerea ◽  
Aspartic Acid ◽  
Mechanism Of Action ◽  
Dna Content ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Lipid Fraction ◽  
Acid Incorporation ◽  
Total Dna

Griseofulvin had no effect on the respiration of Botrytis cinerea but, nevertheless, inhibited growth and caused abnormal hyphal formations, including stunting, spiraling, thickening of the cell wall, and disorientation of growth. Treated cells had an increase in total deoxyribonucleic acid and phosphorus but not in protein, carbohydrate, lipid, and ribonucleic acid. Griseofulvin-treated cells synthesized DNA from labelled glucose and glycine continuously and for much longer periods than control cells so that their total DNA content was greater than that in untreated cells. However, the antibiotic allowed slightly less incorporation with aspartic acid -U-14C as the precursor than the controls. Griseofulvin caused 25 to 50% increased incorporation of carbon into RNA from glucose and glycine but the increases were not due to a prolongation of the synthetic period, and again aspartic acid incorporation was slightly decreased. Griseofulvin was bound to the particulate parts of the cell and was especially high in the lipid fraction of the cell. The antibiotic was not bound to DNA or to RNA.

scholarly journals DEOXYRIBONUCLEIC ACID CYTOPHOTOMETRY OF STAINED HUMAN LEUKOCYTES I. DIFFERENCES AMONG CELL TYPES

1969 ◽  
Vol 17 (4) ◽  
pp. 249-257 ◽  
Author(s):  
BRIAN H. MAYALL
Keyword(s):  
Visible Light ◽  
Dna Content ◽  
Close Agreement ◽  
Deoxyribonucleic Acid ◽  
Cell Types ◽  
Neutrophilic Granulocytes ◽  
Feulgen Reaction ◽  
Human Leukocytes ◽  
Intensity Measurements ◽  
Individual Human

The deoxyribonucleic acid (DNA) content of individual human leukocytes was estimated cytophotometrically using visible light and spreads stained either with gallocyanin-chrome alum following ribonuclease digestion or with the Feulgen reaction. When the cells were measured on a scanning cytophotometer, significant differences in stain intensity were found among slides. Significant differences also were found among the leukocyte types. In gallocyanin-chrome alum preparations, monocytes measured 16% higher than small lymphocytes and 13% higher than neutrophilic granulocytes. In Feulgen preparations, monocytes measured 4% higher than small lymphocytes and 6% higher than neutrophils. These differences among cell types were independent of donor and stain intensity. Measurements of cells within types and within slides frequently showed close agreement, but it is only in this very limited context that the data are consistent with the hypothesis of DNA constancy. Measurements made on a two-wavelength cytophotometer showed a divergence of only 2.1% relative to similar measurements made on the scanning cytophotometer, which suggests that the differences observed among cells and types are unlikely to be artifacts of the instruments. Over-all, the data indicate either that there is variability in DNA content or that DNA is not being expressed correctly by the measured stain content.

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