scholarly journals RNA polymerase activity in bovine spermatozoa.

1977 ◽  
Vol 74 (3) ◽  
pp. 698-706 ◽  
Author(s):  
C D Fuster ◽  
D Farrell ◽  
F A Stern ◽  
N B Hecht
Keyword(s):  
Rna Polymerase ◽  
Sodium Hydroxide ◽  
Ethidium Bromide ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Polymerase Activity ◽  
Intracellular Location ◽  
Bovine Spermatozoa ◽  
Rna Polymerase Activity ◽  
Mature Spermatozoa

Washed mature spermatozoa from bulls incorporate ribonucleoside triphosphates into RNA using an endogenous template. Maximum incorporation was observed at 31 degrees C in the presence of MgCl2, all four ribonucleoside triphosphates, beta-mercaptoethanol, and glycine sodium hydroxide buffer at pH 9.0. The amount of synthesis was linearly dependent upon the concentration of spermatozoa and continued for at least 4 h. Digestion studies revealed the RNA to be present in a protected (intracellular?) location in the spermatozoa. The RNA synthesis was inhibited by ethidium bromide, rifampicin, acriflavine, actinomycin D, and caffeine, but not by alpha-amanitine or rifamycin SV. Fractionation of the spermatozoa by sonication and separation of the heads and tails by centrifugation through a discontinuous gradient revealed that more than half of the total RNA polymerase activity was associated with the tail fraction.

scholarly journals RNA SYNTHESIS BY EXOGENOUS RNA POLYMERASE ON CYTOLOGICAL PREPARATIONS OF CHROMOSOMES

1973 ◽  
Vol 57 (2) ◽  
pp. 538-550 ◽  
Author(s):  
R. Sederoff ◽  
R. Clynes ◽  
M. Poncz ◽  
S. Hachtel
Keyword(s):  
Rna Polymerase ◽  
Dna Sequences ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Polytene Chromosomes ◽  
Polymerase Activity ◽  
Exogenous Rna ◽  
Rna Polymerase Activity ◽  
Chromosome Regions

Cytological preparations of Drosophila polytene chromosomes serve as templates for RNA synthesis carried out by exogenous RNA polymerase (Escherichia coli). Incorporation of labeled ribonucleoside triphosphates into RNA may be observed directly by autoradiography. Because of the effects of rifampicin, actinomycin D, ribonuclease, high salt, and the requirement for all four nucleoside triphosphates, we conclude that the labeling observed over chromosomes is due to DNA-dependent RNA polymerase activity. Using this method, one can observe RNA synthesis in vitro on specific chromosome regions due to the activity of exogenous RNA polymerase. We find that much of the RNA synthesis in this system occurs on DNA sequences which appear to be in a nondenatured state.

scholarly journals RNA Polymerase Activity Assay on Biochips: Correlation between Template DNA Density and RNA Synthesis

2010 ◽  
Vol 31 (7) ◽  
pp. 2107-2109 ◽  
Author(s):  
Bok-Hui Lee ◽  
Hyun-Jung Seo ◽  
So-Hyun Kim ◽  
Woong Jung ◽  
Dong-Woon Kim ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Synthesis ◽  
Polymerase Activity ◽  
Activity Assay ◽  
Rna Polymerase Activity ◽  
Template Dna ◽  
Dna Density

In vitro transcription of RNA in nuclei, nucleoli and chromatin from physarum polycephalum

1977 ◽  
Vol 26 (1) ◽  
pp. 267-279
Author(s):  
K.E. Davies ◽  
I.O. Walker
Keyword(s):  
Rna Polymerase ◽  
Physarum Polycephalum ◽  
Rna Synthesis ◽  
In Vitro Transcription ◽  
Polymerase Activity ◽  
Controlled Conditions ◽  
Rna Polymerase Activity ◽  
Alpha Amanitin

Methods for isolating nuclei, nucleoli and chromatin from Physarum polycephalum which retain high levels of endogenous RNA polymerase activity are described. Under carefully controlled conditions with respect to mono- and divalent cation concentrations RNA synthesis in nuclei displayed linear kinetics for at least 30 min and the RNA products had a similar size distribution to nuclear RNA synthesis observed in vivo. Chromatin showed 60% of the nuclear transcriptional activity but no conditions were found where faithful transcription of the template occurred. Isolated nucleoli were 5-fold more active than nuclei and the endogenous RNA polymerase activity was insensitive to alpha-amanitin. Under carefully controlled conditions, the nucleoli appeared to support the accurate transcription, re-initiation and processing of rRNA chains in vitro.

scholarly journals Mechanism of inhibition of Escherichia coli RNA polymerase by captan

1982 ◽  
Vol 201 (1) ◽  
pp. 145-151 ◽  
Author(s):  
J W Dillwith ◽  
R A Lewis
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Rna Synthesis ◽  
Polymerase Activity ◽  
Mechanism Of Inhibition ◽  
Dna Binding Site ◽  
Rna Polymerase Activity ◽  
Dna Template ◽  
Template Dna

Captan (N-trichloromethylthiocyclohex-4-ene-1,2-dicarboximide) was shown to inhibit RNA synthesis in vitro catalysed by Escherichia coli RNA polymerase. Incorporation of [gamma-32P]ATP and [gamma-32P]GTP was inhibited by captan to the same extent as overall RNA synthesis. The ratio of [3H]UTP incorporation to that of [gamma-32P]ATP or of [gamma-32P]GTP in control and captan-treated samples indicated that initiation was inhibited, but the length of RNA chains being synthesized was not altered by captan treatment. Limited-substrate assays in which re-initiation of RNA chains did not occur also showed that captan had no effect on the elongation reaction. Studies which measured the interaction of RNA polymerase with template DNA revealed that the binding of enzyme to DNA was inhibited by captan. Glycerol-gradient sedimentation of the captan-treated RNA polymerase indicated that the inhibition of the enzyme was irreversible and did not result in dissociation of its subunits. These data are consistent with a mechanism in which RNA polymerase activity was irreversibly altered by captan, resulting in an inability of the enzyme to bind to the template. This interaction was probably at the DNA-binding site on the polymerase and did not involve reaction of captan with the DNA template.

Alterations in myocardial RNA synthesis and RNA polymerase activity during normal growth

1983 ◽  
Vol 61 (4) ◽  
pp. 341-348 ◽  
Author(s):  
James D. McCully ◽  
Paul B. Taylor ◽  
Nancy L. Morrison
Keyword(s):  
Rna Polymerase ◽  
Rna Synthesis ◽  
Specific Activity ◽  
Normal Growth ◽  
Polymerase Activity ◽  
Nuclear Activity ◽  
Chromatin Template ◽  
Male Wistar Rats ◽  
Rna Polymerase Activity ◽  
Age Range

The effect of normal growth (hypertrophy) on myocardial nuclear activity was investigated using male Wistar rats at 21, 50, and 100 days of age. Cardiac mass increased sevenfold during this age range. The concentration of RNA (mg∙g−1) was the highest at 21 days and decreased 48% by 50 days of age and 68% after 100 days of development. RNA synthesis, corrected for alterations in the specific activity of the cytoplasmic nucleotide pool, was the highest at 21 days of age. After 50 days of growth, uridine incorporation was decreased fivefold. With continual growth (100 days), RNA synthesis was still reduced compared with the 21-day animals. RNA polymerase activity in myocyte nuclei showed little change in activity from 21 to 100 days of age. However, in the nonmyocyte fraction, RNA polymerase decreased threefold after 50 days of development. Collectively, these data suggest that the large decrease in myocardial RNA synthesis cannot be accounted for by a change in nuclear RNA polymerase activity and that an alteration in chromatin template capacity may be involved during this form of cardiac growth.

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