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Effect of the exotoxin of Bacillus thuringiensis on normal and ecdysone-stimulated ribonucleic acid polymerase activity in intact nuclei from the fat-body of Sarcophaga bullata larvae.

ABSTRACT: A nuclear preparation from the fat-body of Sarcophaga bullata was obtained which incorporates nucleotides at a steady rate. Two activities, differing in their response to alpha-amanitin, are present. The activities are not separated by changes in the concentration of (NH(4))(2)SO(4) as effectively as in mammalian nuclei. The activity resistant to alpha-amanitin is stimulated by ecdysone, and both normal and the ecdysone-stimulated activities are inhibited by the exotoxin. The amanitin-sensitive enzyme is also inhibited by exotoxin, but higher concentrations are required.

SUBMITTER: Beebee TJ

PROVIDER: S-EPMC1165918 | biostudies-other | 1973 Sep

REPOSITORIES: biostudies-other

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Differential inhibition of mammalian ribonucleic acid polymerases by an exotoxin from Bacillus thuringiensis. The direct observation of nucleoplasmic ribonucleic acid polymerase activity in intact nuclei.

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Inhibition of hepatic deoxyribonucleic acid-dependent ribonucleic acid polymerases by the exotoxin of Bacillus thuringiensis in comparison with the effects of -amanitin and cordycepin.

Project description:The action of Bacillus thuringiensis exotoxin, a structural analogue of ATP, on mouse liver DNA-dependent RNA polymerases was studied and its effects were compared with those of alpha-amanitin and cordycepin. (1) Administration of exotoxin in vivo caused a marked decrease in RNA polymerase activity of isolated nuclei at various concentrations of Mg(2+), Mn(2+) and (NH(4))(2)SO(4). A similar action was recorded after addition of exotoxin to isolated nuclei from control or exotoxin-treated mice. (2) Chromatographic separation of nuclear RNA polymerases from mice treated in vivo with exotoxin showed a drastic decrease of the peak of nucleoplasmic RNA polymerase, whereas the peak of nucleolar RNA polymerase remained unaltered. The same effect was observed after administration of alpha-amanitin in vivo, but cordycepin did not alter the relative amounts of the two main RNA polymerase peaks. (3) Administration of exotoxin in vivo did not alter the template activity of isolated DNA or chromatin tested with different fractions of RNA polymerase from control or exotoxin-treated mice. (4) Addition of exotoxin to isolated liver RNA polymerases inhibited both enzyme fractions. However, the alpha-amanitin-sensitive RNA polymerase was also 50-100-fold more sensitive to exotoxin inhibition than was the alpha-amanitin-insensitive RNA polymerase. Kinetic analysis indicated the exotoxin produces a competitive inhibition with ATP on the nucleolar enzyme, but a mixed type of inhibition with nucleoplasmic enzyme. The results obtained indicate that the B. thuringiensis exotoxin inhibits liver RNA synthesis by affecting nuclear RNA polymerases, showing a preferential inhibition of the nucleoplasmic alpha-amanitin-sensitive RNA polymerase.

| S-EPMC1174051 | biostudies-other

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