Studies on Isolated Brain Nuclear DNA-Dependent RNA Polymerase

1972 ◽  
Vol 50 (3) ◽  
pp. 299-304 ◽  
Author(s):  
Vijendra K. Singh ◽  
S. C. Sung
Keyword(s):  
Enzyme Activity ◽  
Rna Polymerase ◽  
Actinomycin D ◽  
Nuclear Dna ◽  
Divalent Cations ◽  
Polymerase Activity ◽  
Maximum Activity ◽  
Optimum Concentration ◽  
Brain Nuclei ◽  
Rna Polymerase Activity

DNA-dependent RNA polymerase, which was isolated from beef brain nuclei, was stimulated by Mn2+ and Mg2+. The polymerase was about four times as active with Mn2+ as with Mg2+ but both of these divalent cations were required for maximum activity. KCl stimulated the enzyme activity with an optimum concentration around 0.2 M. The stimulation by KCl was much more pronounced in the presence of Mn2+ or Mn2+ plus Mg24 than in the presence of Mg2+ alone. Spermidine and spermine also stimulated the RNA polymerase activity and this stimulation was much greater in the presence of Mn2+ than in the presence of Mg2+ or both together. With Mn2+ plus Mg2+, spermidine had little or no stimulatory effect when heat-denatured DNA served as template, whereas with Mn2+ alone, spermidine markedly stimulated the enzyme in the presence of either native or heat-denatured DNA. The effect of spermidine was different with Mg2+ alone. The enzyme was inhibited as much as 75% by actinomycin D and almost completely by α-amanitin but not by rifampicin. Yeast RNA inhibited the enzyme activity considerably while spermidine appeared to overcome and prevent this inhibition of RNA polymerase.

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 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 Effect of Phenylhydrazine-induced Hemolytic Anemia on Nuclear RNA Polymerase Activity of the Mouse Spleen

Blood ◽  
1973 ◽  
Vol 42 (2) ◽  
pp. 257-266 ◽  
Author(s):  
Jerry L. Spivak ◽  
Dennis Toretti ◽  
Herbert W. Dickerman
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Hemolytic Anemia ◽  
Nuclear Dna ◽  
Polymerase Activity ◽  
Tenfold Increase ◽  
Rna Polymerase Activity ◽  
Low Ionic Strength ◽  
Dna Template ◽  
Nuclear Rna Polymerase

Abstract The DNA-dependent RNA polymerase of nuclei from lymphoid-rich and erythroid-rich mouse spleens was compared in regard to requirements and conditions of the reactions. The inhibition by either actinomycin D or pancreatic DNase indicated that a DNA template was required for the observed reactions. Following the induction of a hemolytic anemia by the administration of phenylhydrazine, there was a tenfold increase in the nuclear polymerase activity per milligram nuclear DNA of the developing erythropoietic spleen when the assays were done at low ionic strength and more than a threefold increase at high ionic strength. The peak rise in polymerase activity precedes the maximal development of the erythropoietic spleen by 3 days.

Ribonucleic Acid Polymerase Induced in L-Cells Infected with Vesicular Stomatitis Virus

1973 ◽  
Vol 51 (5) ◽  
pp. 721-729 ◽  
Author(s):  
Helene Galet ◽  
Joseph G. Shedlarski Jr. ◽  
Ludvik Prevec
Keyword(s):  
Enzyme Activity ◽  
Rna Polymerase ◽  
Vesicular Stomatitis Virus ◽  
Ribonucleic Acid ◽  
Polymerase Activity ◽  
L Cells ◽  
Ribonucleoprotein Complexes ◽  
Rna Polymerase Activity ◽  
Vesicular Stomatitis ◽  
Long Chain

RNA polymerase activity capable of synthesizing long-chain, heteropolymeric RNA is associated with specific ribonucleoprotein complexes present in the cytoplasm of cells infected with vesicular stomatitis virus (VSV). The synthesis is independent of added exogenous template and the product labelled with GTP precursors can be totally annealed with excess virion RNA. The enzyme activity is therefore a transcriptase similar to that observed in association with the mature VSV virion.

AN ASSAY FOR RNA POLYMERASE ACTIVITY IN RAT BRAIN NUCLEI: EFFECTS OF INJECTION OF OESTRADIOL BENZOATE

1980 ◽  
Vol 85 (2) ◽  
pp. 341-347 ◽  
Author(s):  
B. D. GREENSTEIN
Keyword(s):  
Rna Polymerase ◽  
Rat Brain ◽  
Preliminary Evidence ◽  
Polymerase Activity ◽  
Female Rats ◽  
Oestrogen Receptors ◽  
Guanosine Triphosphate ◽  
Brain Nuclei ◽  
Oestradiol Benzoate ◽  
Rna Polymerase Activity

A sensitive, reliable assay of activity of RNA polymerase in purified nuclei from the rat brain is described. The assay measured the incorporation of [3H]guanosine triphosphate into RNA by nuclei. Little incorporation occurred in the presence of α-amanitin (400 ng) suggesting that RNA polymerase B activity was being measured. Preliminary evidence showed that after administration of oestradiol benzoate to 18-day-old female rats RNA polymerase activity was raised in areas of brain known to contain oestrogen receptors.

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