Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Apr;73(4):1029–1033. doi: 10.1073/pnas.73.4.1029

Transcription in yeast: alpha-amanitin sensitivity and other properties which distinguish between RNA polymerases I and III.

L D Schultz, B D Hall
PMCID: PMC430193  PMID: 772676

Abstract

Three peaks of DNA-dependent RNA polymerase (RNA nucleotidyltransferase) activity are resolved by chromatography of a sonicated yeast cell extract on DEAE-Sephadex. The enzymes, which are named RNA polymerases I, II, and III in order of elution, show similar catalytic properties to the vertebrate class I, class II, and class III RNA polymerases, respectively. Yeast RNA polymerase III is readily distinguished from yeast polymerase I by its biphasic amnonium sulfate activation profile with native DNA templates, greater enzymatic activity with poly[d(I-C)] than with native salmon sperm DNA, and distinctive chromatographic elution positions from DEAE-cellulose (0.12 M ammonium sulfate) compared with DEAE-Sephadex (0.32 M ammonium sulfate). The three yeast RNA polymerases also show significant differences in alpha-amanitin inhibition. RNA polymerase II is the most sensitive (50% inhibition at 1.0 mug of alpha-amanitin per ml). Contrary to the results for vertebrate systems, yeast polymerase I can be completely inhibited by alpha-amanitin at high concentrations (50% inhibition at 600 mug/ml) while yeast RNA polymerase II BEGINS TO SHOW SIGNIFICANT INHIBITION ONLY AT CONCENTRATIONS EXCEEDING 1 MG/ML. Therefore, yeast RNA polymerases I and III show a pattern of alpha-amanitin sensitivity that is the reverse of that seen for the analogous vertebrate RNA polymerases.

Full text

PDF
1029

Selected References

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

  1. Adman R., Schultz L. D., Hall B. D. Transcription in yeast: separation and properties of multiple FNA polymerases. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1702–1706. doi: 10.1073/pnas.69.7.1702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Austoker J. L., Beebee T. J., Chesterton C. J., Butterworth P. H. DNA-dependent RNA polymerase activity of Chinese hamster kidney cells sensitive to high concentrations of alpha-amanitin. Cell. 1974 Nov;3(3):227–234. doi: 10.1016/0092-8674(74)90136-6. [DOI] [PubMed] [Google Scholar]
  3. Beebee T. J., Butterworth P. H. Transcription specificity of Xenopus laevis RNA polymerase A. FEBS Lett. 1974 Oct 15;47(2):304–306. doi: 10.1016/0014-5793(74)81035-5. [DOI] [PubMed] [Google Scholar]
  4. Brogt T. M., Planta R. J. Characteristics of DNA-dependent RNA polymerase activity from isolated yeast nuclei. FEBS Lett. 1972 Jan 15;20(1):47–52. doi: 10.1016/0014-5793(72)80014-0. [DOI] [PubMed] [Google Scholar]
  5. Buhler J. M., Sentenac A., Fromageot P. Isolation, structure, and general properties of yeast ribonucleic acid polymerase A (or I). J Biol Chem. 1974 Sep 25;249(18):5963–5970. [PubMed] [Google Scholar]
  6. Chambon P. Eukaryotic nuclear RNA polymerases. Annu Rev Biochem. 1975;44:613–638. doi: 10.1146/annurev.bi.44.070175.003145. [DOI] [PubMed] [Google Scholar]
  7. Dezelee S., Sentenac A., Fromageot P. Role of DNA-RNA hybrids in eukaryots 1. Purification of yeast RNA polymerase B. FEBS Lett. 1972 Mar;21(1):1–6. doi: 10.1016/0014-5793(72)80148-0. [DOI] [PubMed] [Google Scholar]
  8. Dezélée S., Sentenac A. Role of DNA-RNA hybrids in eukaryotes. Purification and properties of yeast RNA polymerase B. Eur J Biochem. 1973 Apr 2;34(1):41–52. doi: 10.1111/j.1432-1033.1973.tb02726.x. [DOI] [PubMed] [Google Scholar]
  9. Hartwell L. H. Macromolecule synthesis in temperature-sensitive mutants of yeast. J Bacteriol. 1967 May;93(5):1662–1670. doi: 10.1128/jb.93.5.1662-1670.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lohr D., Van Holde K. E. Yeast chromatin subunit structure. Science. 1975 Apr 11;188(4184):165–166. doi: 10.1126/science.1090006. [DOI] [PubMed] [Google Scholar]
  11. Ponta H., Ponta U., Wintersberger E. DNA-dependent RNA polymerases from yeast. Partial characterization of three nuclear enzyme activities. FEBS Lett. 1971 Nov 1;18(2):204–208. doi: 10.1016/0014-5793(71)80445-3. [DOI] [PubMed] [Google Scholar]
  12. Ponta H., Ponta U., Wintersberger E. Purification and properties of DNA-dependent RNA polymerases from yeast. Eur J Biochem. 1972 Aug 18;29(1):110–118. doi: 10.1111/j.1432-1033.1972.tb01964.x. [DOI] [PubMed] [Google Scholar]
  13. Roeder R. G. Multiple forms of deoxyribonucleic acid-dependent ribonucleic acid polymerase in Xenopus laevis. Isolation and partial characterization. J Biol Chem. 1974 Jan 10;249(1):241–248. [PubMed] [Google Scholar]
  14. 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]
  15. Schwartz L. B., Sklar V. E., Jaehning J. A., Weinmann R., Roeder R. G. Isolation and partial characterization of the multiple forms of deoxyribonucleic acid-dependent ribonucleic acid polymerase in the mouse myeloma, MOPC 315. J Biol Chem. 1974 Sep 25;249(18):5889–5897. [PubMed] [Google Scholar]
  16. Sebastian J., Bhargava M. M., Halvorson H. O. Nuclear deoxyribonucleic acid-dependent ribonucleic acid polymerases from Saccharomyces cerevisiae. J Bacteriol. 1973 Apr;114(1):1–6. doi: 10.1128/jb.114.1.1-6.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sergeant A., Krsmanovic V. KB cell RNA polymerases: occurrence of nucleoplasmic enzyme 3. FEBS Lett. 1973 Sep 15;35(2):331–335. doi: 10.1016/0014-5793(73)80316-3. [DOI] [PubMed] [Google Scholar]
  18. Sklar V. E., Schwartz L. B., Roeder R. G. Distinct molecular structures of nuclear class I, II, and III DNA-dependent RNA polymerases. Proc Natl Acad Sci U S A. 1975 Jan;72(1):348–352. doi: 10.1073/pnas.72.1.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tipper D. J. Inhibition of yeast ribonucleic acid polymerases by thiolutin. J Bacteriol. 1973 Oct;116(1):245–256. doi: 10.1128/jb.116.1.245-256.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Valenzuela P., Hager G. L., Weinberg F., Rutter W. J. Molecular structure of yeast RNA polymerase III: demonstration of the tripartite transcriptive system in lower eukaryotes. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1024–1028. doi: 10.1073/pnas.73.4.1024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weil P. A., Blatti S. P. Partial purification and properties of calf thymus deoxyribonucleic acid dependent RNA polymerase III. Biochemistry. 1975 Apr 22;14(8):1636–1642. doi: 10.1021/bi00679a015. [DOI] [PubMed] [Google Scholar]
  22. Weinmann R., Raskas H. J., Roeder R. G. Role of DNA-dependent RNA polymerases II and III in transcription of the adenovirus genome late in productive infection. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3426–3439. doi: 10.1073/pnas.71.9.3426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weinmann R., Roeder R. G. Role of DNA-dependent RNA polymerase 3 in the transcription of the tRNA and 5S RNA genes. Proc Natl Acad Sci U S A. 1974 May;71(5):1790–1794. doi: 10.1073/pnas.71.5.1790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wintersberger U., Smith P., Letnansky K. Yeast chromatin. Preparation from isolated nuclei, histone composition and transcription capacity. Eur J Biochem. 1973 Feb 15;33(1):123–130. doi: 10.1111/j.1432-1033.1973.tb02663.x. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES