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

Stirpe, Fiorenzo; Fiume, L.

doi:10.1042/bj1050779

Cited by 187 publications

(44 citation statements)

References 16 publications

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“…We have also * A. Rifkind and P. Gillette, personal communications. established that of the two RNA polymerases identifiable in chick embryo liver nuclei [ 131 it is the Mn2+-(NH4)2 SO4 stimulated enzyme which is sensitive to this toxin, a finding which conforms to the results of other workers [7,8,141. These findings indicate that the avian embryo is another experimental species in which enzyme induction elicited by steroids [3,121 can be shown to be sensitive to the inhibitory action of a-amanitin.…”

Section: Resultssupporting

confidence: 85%

Inhibitory effect of a‐amanitin on the induction of δ‐aminolevulinate synthetase in chick embryo liver

Incefy

Kappas

1971

FEBS Letters

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Section: Resultssupporting

confidence: 85%

Inhibitory effect of a‐amanitin on the induction of δ‐aminolevulinate synthetase in chick embryo liver

Incefy

Kappas

1971

FEBS Letters

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“…RNA was determined by absorption at 260 nm of the material that precipitated in cold 0.50 N perchloric acid and dissolved after alkaline hydrolysis (1 N NaOH, 370, 18 hr), with 25 Nucleoli were separated from nucleoplasm by the method of Penman (10). For sedimentation analysis of RNA synthesized in ghost monolayers, the RNA was extracted by the phenolchloroform method (10) and centrifuged through a 15-30% sucrose gradient containing 10 mM Tris * HCl (pH 7.2)-0.10 M NaCl-1 mM EDTA-0.5% Na dodecyl sulfate, at 20,000 rpm for 15 hr in an SW27 rotor at 230.…”

Section: Methodsmentioning

confidence: 99%

Regulation of RNA Synthesis in Fibroblasts During Transition from Resting to Growing State

Mauck

Green

1973

Proc. Natl. Acad. Sci. U.S.A.

111

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Addition of serum, containing fibroblast growth factors, to a culture of resting 3T6 cells stimulates a transition to the growing state. Studies of ghost monolayers prepared with the aid of detergent at intervals after stimulation showed an increase in the rate of ribosomal RNA synthesis within 10 min. The rate continued to increase for many hours and reached a level 2.5-to 3.5-fold higher by the time DNA synthesis began. The increasing rate of ribosomal RNA synthesis appeared independent of an increase in the number of ribosomal genes, since it was not affected by prevention of DNA synthesis with cytosine arabinoside.In contrast to ribosomal RNA, the overall rate of transscription of heterogeneous nuclear RNA was not directly affected by serum growth factors and does not appear to be regulated during the transition from resting to growing state. It seems, instead, to be fixed in relation to the amount of template, for it increases proportionally to DNA content.The rate of RNA synthesis in mouse-fibroblast line 3T6 can be measured as the incorporation of labeled ribonucleoside triphosphates by nonviable "ghost monolayers" prepared with the aid of the detergent, NP-40 (1). Estimates of the rate of RNA synthesis under these conditions are not affected by the cell-membrane permeability barrier and the slowly equilibrating cell-nucleotide pools, which complicate interpretation of incorporation rates in whole cells (2)(3)(4)(5)(6).After stimulation of resting cultures of 3T6 with serum containing fibroblast growth factors, an increase in total RNA synthesis could be detected in ghost monolayers prepared less than 30 min later (1). The rate rose 2-fold by the time DNA synthesis began, and continued to rise for at least 20 hr. No attempt was made to distinguish effects on transcription of different classes of RNA.We describe here experiments on the rates of synthesis of ribosomal RNA (rRNA) and of heterogeneous nuclear RNA in ghost monolayers prepared during the serum-induced transition from resting to growing state. These two classes of RNA respond quite differently, indicating that their synthesis is controlled independently. An increased rate of ribosome synthesis is lprobably an essential part of the preparation for DNA synthesis, but the overall synthesis of heterogeneous nuclear RNA, the precursor of cytoplasmic mRNA (7) Assay of RNA Synthesis in Ghost Monolayers. All operations were done at 37°. The medium was removed and the monolayer was washed with 2 ml of serum-free medium. The monolayer was then treated with 1 ml of 0.5% NP-40 in assay buffer containing 0.05 M Tris HCl (pH 7.8), 5 mM MgCJ2, 6 mM KCl, 75 mM (NH4)2SO4, 0.20 M sucrose, and 1 mM freshly prepared dithiothreitol. After exactly 3 min, the NP-40 solution was removed and the ghost monolayer was washed with assay buffer. 0.80 ml of a solution of [3H]UTP (10 ,4Ci, [35][36][37][38][39][40][41][42][43][44][45][46][47][48][49][50] Ci/mmol), 5 mM ATP, 0.2 mM CTP, and 0.2 mM GTP in assay buffer was gently pipetted onto the ghost monolayer. The reac...

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“…However, it remained possible that the reaction of unconventional splicing was coupled with transcription, and that nuclear XBP1 mRNA was spliced in the nucleus and rapidly transported to the cytoplasm. To exclude this possibility, de novo transcription was halted by adding α-amanitin, an inhibitor of RNA polymerase II (Stirpe and Fiume, 1967) (Fig. 6A-C).…”

Section: Xbp1(u) Mrna Is Localized In the Cytoplasm And Associated Wimentioning

confidence: 99%

Unconventional splicing ofXBP1mRNA occurs in the cytoplasm during the mammalian unfolded protein response

Uemura

Oku

Mori

et al. 2009

Journal of Cell Science

135

104

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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

Cited by 187 publications

References 16 publications

Inhibitory effect of a‐amanitin on the induction of δ‐aminolevulinate synthetase in chick embryo liver

Inhibitory effect of a‐amanitin on the induction of δ‐aminolevulinate synthetase in chick embryo liver

Regulation of RNA Synthesis in Fibroblasts During Transition from Resting to Growing State

Unconventional splicing ofXBP1mRNA occurs in the cytoplasm during the mammalian unfolded protein response

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