Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1972 May 1;53(2):523–531. doi: 10.1083/jcb.53.2.523

EFFECTS OF α-AMANITIN ON IN VITRO LABELING OF RNA FROM DEFINED NUCLEAR COMPONENTS IN SALIVARY GLAND CELLS FROM CHIRONOMUS TENTANS

E Egyházi 1, B D'Monte 1, J-E Edström 1
PMCID: PMC2108739  PMID: 5025109

Abstract

The effect of α-amanitin on nucleoside labeling of RNA in nucleoli, chromosomes, nuclear sap, and cytoplasm from Chironomus tentans salivary gland cells was investigated by radioautography and gel electrophoresis. Preribosomal RNA formation and processing in the nucleolus was not measurably influenced by the drug, and both 28 S and 18 S ribosomal RNA were transferred to the cytoplasm. In the chromosomes the heterogeneous RNA labeling was completely inhibited for the large size range (above 45–50 S) and partially for the low range. The labeling of 4–5 S chromosomal RNA was only moderately reduced. Most of the chromosomes showed radioautographically a disappearance of the normal band pattern, but some retained a pattern of weakly labeled bands. The electrophoretic results for the nuclear sap paralleled those for the chromosomes. The effect of α-amanitin on RNA labeling in these cells is similar but not identical to that of the substituted benzimidazole 5,6-dichloro-1(β-D-ribofuranosyl) benzimidazole (DRB).

Full Text

The Full Text of this article is available as a PDF (593.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beermann W. Effect of -amantine on puffing and intranuclear RNA synthesis in Chironomus salivary glands. Chromosoma. 1971;34(2):152–167. doi: 10.1007/BF00285183. [DOI] [PubMed] [Google Scholar]
  2. CANNON G. B. CULTURE OF INSECT SALIVARY GLANDS IN A CHEMICALLY DEFINED MEDIUM. Science. 1964 Nov 20;146(3647):1063–1063. doi: 10.1126/science.146.3647.1063. [DOI] [PubMed] [Google Scholar]
  3. Daneholt B., Edström J. E., Egyházi E., Lambert B., Ringborg U. Chromosomal RNA synthesis in polytene chromosomes of Chironomus tentans. Chromosoma. 1969;28(4):399–417. doi: 10.1007/BF00284257. [DOI] [PubMed] [Google Scholar]
  4. Edström J. E., Daneholt B. Sedimentation properties of the newly synthesized RNA from isolated nuclear components of Chironomus tentans salivary gland cells. J Mol Biol. 1967 Sep 14;28(2):331–343. doi: 10.1016/s0022-2836(67)80013-5. [DOI] [PubMed] [Google Scholar]
  5. Edström J. E., Egyházi E., Daneholt B., Lambert B., Ringborg U. Localization of newly synthesized low molecular weight RNA in polytene chromosomes from Chironomus tentans. Chromosoma. 1971;35(4):431–442. doi: 10.1007/BF02451449. [DOI] [PubMed] [Google Scholar]
  6. Egyházi E., Daneholt B., Edström J. E., Lambert B., Ringborg U. Low molecular weight RNA in cell components of Chironomus tentans salivary glands. J Mol Biol. 1969 Sep 28;44(3):517–532. doi: 10.1016/0022-2836(69)90377-5. [DOI] [PubMed] [Google Scholar]
  7. Jacob S. T., Muecke W., Sajdel E. M., Munro H. N. Evidence for extranucleolar control of RNA synthesis in the nucleolus. Biochem Biophys Res Commun. 1970 Jul 27;40(2):334–342. doi: 10.1016/0006-291x(70)91014-4. [DOI] [PubMed] [Google Scholar]
  8. Jacob S. T., Sajdel E. M., Munro H. N. Different responses of soluble whole nuclear RNA polymerase and soluble nucleolar RNA polymerase to divalent cations and to inhibition by alpha-amanitin. Biochem Biophys Res Commun. 1970 Feb 20;38(4):765–770. doi: 10.1016/0006-291x(70)90647-9. [DOI] [PubMed] [Google Scholar]
  9. Lindell T. J., Weinberg F., Morris P. W., Roeder R. G., Rutter W. J. Specific inhibition of nuclear RNA polymerase II by alpha-amanitin. Science. 1970 Oct 23;170(3956):447–449. doi: 10.1126/science.170.3956.447. [DOI] [PubMed] [Google Scholar]
  10. Niessing J., Schnieders B., Kunz W., Seifart K. H., Sekeris C. E. Inhibition of RNA synthesis by alpha-amanitin in vivo. Z Naturforsch B. 1970 Oct;25(10):1119–1125. doi: 10.1515/znb-1970-1011. [DOI] [PubMed] [Google Scholar]
  11. Novello F., Stirpe F. Experimental conditions affecting ribonucleic acid polymerase in isolated rat liver nuclei. Effect of nucleoside triphosphate concentration, temperature, ammonium sulphate and heparin. Biochem J. 1969 May;112(5):721–727. doi: 10.1042/bj1120721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ringborg U., Daneholt B., Edström J. E., Egyházi E., Lambert B. Electrophoretic characterization of nucleolar RNA from Chironomus tentans salivary gland cells. J Mol Biol. 1970 Jul 28;51(2):327–340. doi: 10.1016/0022-2836(70)90146-4. [DOI] [PubMed] [Google Scholar]
  13. Ringborg U., Daneholt B., Edström J. E., Egyházi E., Rydlander L. Evidence for transport of preribosomal RNA from the nucleolus to the chromosomes in Chironomus tentans salivary gland cells. J Mol Biol. 1970 Aug;51(3):679–686. doi: 10.1016/0022-2836(70)90016-1. [DOI] [PubMed] [Google Scholar]
  14. Ringborg U., Rydlander L. Nucleolar-derived ribonucleic acid in chromosomes, nuclear sap, and cytoplasm of Chironomus tentans salivary gland cells. J Cell Biol. 1971 Nov;51(21):355–368. doi: 10.1083/jcb.51.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ritossa F. M., Atwood K. C., Spiegelman S. On the redundancy of DNA complementary to amino acid tranfer RNA and its absence from the nucleolar organizer region of Drosophila melanogaster. Genetics. 1966 Aug;54(2):663–676. doi: 10.1093/genetics/54.2.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roeder R. G., Rutter W. J. Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969 Oct 18;224(5216):234–237. doi: 10.1038/224234a0. [DOI] [PubMed] [Google Scholar]
  17. Roeder R. G., Rutter W. J. Specific nucleolar and nucleoplasmic RNA polymerases. Proc Natl Acad Sci U S A. 1970 Mar;65(3):675–682. doi: 10.1073/pnas.65.3.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Seifart K. H., Sekeris C. E. Alpha-amanitin, a specific inhibitor of transcription by mammalian RNA-polymerase. Z Naturforsch B. 1969 Dec;24(12):1538–1544. doi: 10.1515/znb-1969-1211. [DOI] [PubMed] [Google Scholar]
  19. Stirpe F., Fiume L. Studies on the pathogenesis of liver necrosis by alpha-amanitin. Effect of alpha-amanitin on ribonucleic acid synthesis and on ribonucleic acid polymerase in mouse liver nuclei. Biochem J. 1967 Nov;105(2):779–782. doi: 10.1042/bj1050779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tartof K. D., Perry R. P. The 5 s RNA genes of Drosophila melanogaster. J Mol Biol. 1970 Jul 28;51(2):171–183. doi: 10.1016/0022-2836(70)90135-x. [DOI] [PubMed] [Google Scholar]
  21. Wieland T. Poisonous principles of mushrooms of the genus Amanita. Four-carbon amines acting on the central nervous system and cell-destroying cyclic peptides are produced. Science. 1968 Mar 1;159(3818):946–952. doi: 10.1126/science.159.3818.946. [DOI] [PubMed] [Google Scholar]
  22. Wimber D. E., Steffensen D. M. Localization of 5S RNA genes on Drosophila chromosomes by RNA-DNA hybridization. Science. 1970 Nov 6;170(3958):639–641. doi: 10.1126/science.170.3958.639. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES