scholarly journals Studies on the pathogenesis of liver necrosis by α-amanitin. Effect of α-amanitin on ribonucleic acid synthesis and on ribonucleic acid polymerase in mouse liver nuclei

1967 ◽  
Vol 105 (2) ◽  
pp. 779-782 ◽  
Author(s):  
F. Stirpe ◽  
L. Fiume

1. Injection of α-amanitin to mice causes a decreased incorporation of [6−14C]-orotic acid into liver RNA in vivo. 2. The activity of RNA polymerase activated by Mn2+ and ammonium sulphate is greatly impaired in liver nuclei isolated from mice poisoned with α-amanitin, and is inhibited by the addition of the same toxin in vitro. 3. The activity of the Mg2+-activated RNA polymerase is only slightly affected by α-amanitin either administered to mice or added in vitro.

1970 ◽  
Vol 116 (2) ◽  
pp. 177-180 ◽  
Author(s):  
F. Novello ◽  
L. Fiume ◽  
F. Stirpe

1. α-Amanitin inhibits in vitro the RNA polymerase solubilized from isolated rat liver nuclei. 2. In contrast with previous observations with whole nuclei, the inhibition occurs approximately to the same extent in the presence and in the absence of ammonium sulphate. 3. Evidence is presented that the toxin acts by interacting with the enzyme itself and not with DNA or other components.


1970 ◽  
Vol 120 (2) ◽  
pp. 381-384 ◽  
Author(s):  
D. Rickwood ◽  
H. G. Klemperer

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.


1978 ◽  
Vol 176 (3) ◽  
pp. 715-725 ◽  
Author(s):  
T J C Beebee

1. A nucleoplasmic fraction rich in endogenous RNA polymerase II activity was isolated from rat liver nuclei and conditions were determined under which elongation of RNA molecules initiated in vivo continued at maximal rates in vitro. 2. Elongation rates in vitro were calculated to be about 0.25 nucleotide/s and there were about 7 × 10(3) RNA molecules in the process of being elongated by form-II RNA polymerase per original nucleus. 3. Evidence was obtained suggesting that transcription-dependent release of RNA polymerase II molecules from the template occurred during the incubations in vitro. 4. The nascent RNA was tightly associated with protein and banded as ribonucleoprotein in caesium salt gradients. 5. RNA molecules labelled in vitro were up to 13000 nucleotides in length, but consisted of long unlabelled chains transcribed in vivo with only short labelled sequences added in vitro, and without significant polyadenylation. 6. Hybridization of transcripts in the presence of a vast excess of DNA demonstrated that both form-II RNA polymerase and another enzyme, resistant to low alpha-amanitin concentrations, were synthesizing RNA molecules complementary to both reiterated and unique DNA sequences in the genome.


1970 ◽  
Vol 117 (5) ◽  
pp. 853-859 ◽  
Author(s):  
T. C. Hall ◽  
K. L. Tao

1. A procedure for measuring rates of aminoacyl-tRNA synthesis in vitro and in intact leaves is presented. 2. Leaf discs showed rates close to those of intact leaves. 3. Cell-free preparations showed similar rates when assayed by pyrophosphate exchange, but actual aminoacyl-tRNA formation rates appeared to be much lower. Evidence is presented that dilution of supplied labelled amino acids was a major factor causing the low apparent rates. 4. Attempts to strip endogenous amino acids from plant tRNA resulted in low acceptor capability of the tRNA.


1960 ◽  
Vol 15 (7) ◽  
pp. 453-460 ◽  
Author(s):  
Christoph Scholtissek ◽  
Van R. Potter

1. The incubation conditions for isolated rat liver nuclei are studied by means of nucleic acid determinations and observations under the light microscope.2. A part of the nuclear ribonucleic acid (RNS) labeled in vivo is transferred from the nuclei to the surrounding medium during incubation in vitro. This RNS appears in the postmicrosomal and cell sap fraction. The transfer process is performed without degradation to dialysable products.3. The transferable RNS amounts to about 15% of the total nuclear RNS. It is synthesized in the nucleus rapidly and disappears from there rapidly. Its specific activity surpasses that of the remaining RNS in the nuclei.4. The rapidly labeled RNS is labeled throughout the whole molecule and is degraded quickly during the incubation in vitro.


1969 ◽  
Vol 111 (1) ◽  
pp. 115-119 ◽  
Author(s):  
F. Novello ◽  
F. Stirpe

1. The effects of various ions on the Mg2+- and Mn2+/ammonium sulphate-activated RNA polymerase activities of isolated liver nuclei were studied. 2. The Mg2+-activated RNA polymerase reaction was inhibited by more than 60% by Cd2+, SeO32−, Be2+, Cu2+, Co2+, Ca2+ and La3+, all at 1mm concentrations. 3. The Mn2+/ammonium sulphate-activated RNA polymerase reaction was strongly inhibited by Hg2+, Cd2+, Cu2+ and Ag+. The effect of Hg2+, Cd2+ and Ag+ was relieved by cysteine or mercaptoethanol. 4. Inhibition by Cu2+ was not affected by addition of DNA, and was relieved only partially by EDTA or histidine. 5. No changes of RNA polymerase activities were observed in nuclei isolated from the liver of rats treated with copper albuminate.


1973 ◽  
Vol 132 (4) ◽  
pp. 813-819 ◽  
Author(s):  
J. Austoker ◽  
D. Cox ◽  
A. P. Mathias

1. Radioactive orotic acid, uridine and adenosine were administered to rats by intracisternal injection. The effects of the size of the dose, the specific radioactivity and time on the incorporation into the RNA of unfractionated nuclei of brain tissue were examined to establish appropriate conditions for studies of the relative activities in vivo of the various sorts of brain nuclei fractionated by zonal centrifugation. Uridine is incorporated more efficiently than either orotic acid or adenosine. 2. With [3H]uridine as precursor the astrocytes in zone (II) contain the highest radioactivity except at the beginning of the experiment when the neuronal nuclei of zone (I) are more highly labelled. This fraction utilizes [14C]orotic acid more readily than the nuclei of zone (II). The astrocytic nuclei of zone (III) show a general resemblance to those of zone (II). Considerable differences between the incorporations into the two types of oligodendrocyte nuclei in zones (IV) and (V) are observed. 3. The relative synthetic activities of the major types of brain nuclei in vitro and in vivo are discussed.


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