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. 1987 Feb 25;15(4):1493–1506. doi: 10.1093/nar/15.4.1493

Novobiocin inhibits interactions required for yeast TFIIIB sequestration during stable transcription complex formation in vitro.

S J Felts, P A Weil, R Chalkley
PMCID: PMC340563  PMID: 3547336

Abstract

Novobiocin concentrations normally used to inhibit a putative eukaryotic DNA gyrase have been found to inhibit transcription of a yeast 5S rRNA gene using an in vitro yeast transcription system. Purified RNA polymerase III and three yeast transcription factors (chromatographically separated, partially purified and free of any detectable gyrase activity) were used. Novobiocin prevents specific transcription if added to the in vitro system immediately prior to the addition of transcription factors and RNA polymerase. If a stable transcription factor complex is allowed to form prior to the addition of novobiocin, concentrations of novobiocin as high as 1000 micrograms/ml have no effect on in vitro transcription. Transcription factors TFIIIA and TFIIIC are able to be stably sequestered onto 5SrDNA-cellulose, but factor TFIIIB is not able to associate with the 5SrDNA-TFIIIA-TFIIIC complex in the presence of novobiocin. Although novobiocin is able to precipitate other basic proteins, it does not appear to precipitate any of these class III gene transcription factors, but instead appears to act by disrupting specific factor-factor interactions.

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Selected References

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  1. Bieker J. J., Martin P. L., Roeder R. G. Formation of a rate-limiting intermediate in 5S RNA gene transcription. Cell. 1985 Jan;40(1):119–127. doi: 10.1016/0092-8674(85)90315-0. [DOI] [PubMed] [Google Scholar]
  2. Bieker J. J., Roeder R. G. Characterization of the nucleotide requirement for elimination of the rate-limiting step in 5 S RNA gene transcription. J Biol Chem. 1986 Jul 25;261(21):9732–9738. [PubMed] [Google Scholar]
  3. Bogenhagen D. F., Wormington W. M., Brown D. D. Stable transcription complexes of Xenopus 5S RNA genes: a means to maintain the differentiated state. Cell. 1982 Feb;28(2):413–421. doi: 10.1016/0092-8674(82)90359-2. [DOI] [PubMed] [Google Scholar]
  4. Carey M. F., Gerrard S. P., Cozzarelli N. R. Analysis of RNA polymerase III transcription complexes by gel filtration. J Biol Chem. 1986 Mar 25;261(9):4309–4317. [PubMed] [Google Scholar]
  5. Cotten M., Bresnahan D., Thompson S., Sealy L., Chalkley R. Novobiocin precipitates histones at concentrations normally used to inhibit eukaryotic type II topoisomerase. Nucleic Acids Res. 1986 May 12;14(9):3671–3686. doi: 10.1093/nar/14.9.3671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Giacherio D., Hager L. P. A poly(dT)-stimulated ATPase activity associated with simian virus 40 large T antigen. J Biol Chem. 1979 Sep 10;254(17):8113–8116. [PubMed] [Google Scholar]
  7. Gottesfeld J. M. Novobiocin inhibits RNA polymerase III transcription in vitro by a mechanism distinct from DNA topoisomerase II. Nucleic Acids Res. 1986 Mar 11;14(5):2075–2088. doi: 10.1093/nar/14.5.2075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hazuda D. J., Wu C. W. DNA-activated ATPase activity associated with Xenopus transcription factor A. J Biol Chem. 1986 Sep 15;261(26):12202–12208. [PubMed] [Google Scholar]
  9. Klekamp M. S., Weil P. A. Partial purification and characterization of the Saccharomyces cerevisiae transcription factor TFIIIB. J Biol Chem. 1986 Feb 25;261(6):2819–2827. [PubMed] [Google Scholar]
  10. Klekamp M. S., Weil P. A. Specific transcription of homologous class III genes in yeast-soluble cell-free extracts. J Biol Chem. 1982 Jul 25;257(14):8432–8441. [PubMed] [Google Scholar]
  11. Kmiec E. B., Ryoji M., Worcel A. Gyration is required for 5S RNA transcription from a chromatin template. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1305–1309. doi: 10.1073/pnas.83.5.1305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kmiec E. B., Worcel A. The positive transcription factor of the 5S RNA gene induces a 5S DNA-specific gyration in Xenopus oocyte extracts. Cell. 1985 Jul;41(3):945–953. doi: 10.1016/s0092-8674(85)80075-1. [DOI] [PubMed] [Google Scholar]
  13. Mizuuchi K., O'Dea M. H., Gellert M. DNA gyrase: subunit structure and ATPase activity of the purified enzyme. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5960–5963. doi: 10.1073/pnas.75.12.5960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ruet A., Camier S., Smagowicz W., Sentenac A., Fromageot P. Isolation of a class C transcription factor which forms a stable complex with tRNA genes. EMBO J. 1984 Feb;3(2):343–350. doi: 10.1002/j.1460-2075.1984.tb01809.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ryoji M., Worcel A. Chromatin assembly in Xenopus oocytes: in vivo studies. Cell. 1984 May;37(1):21–32. doi: 10.1016/0092-8674(84)90297-6. [DOI] [PubMed] [Google Scholar]
  16. Sealy L., Cotten M., Chalkley R. Novobiocin inhibits passive chromatin assembly in vitro. EMBO J. 1986 Dec 1;5(12):3305–3311. doi: 10.1002/j.1460-2075.1986.tb04644.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Segall J. Assembly of a yeast 5 S RNA gene transcription complex. J Biol Chem. 1986 Sep 5;261(25):11578–11584. [PubMed] [Google Scholar]
  18. Setzer D. R., Brown D. D. Formation and stability of the 5 S RNA transcription complex. J Biol Chem. 1985 Feb 25;260(4):2483–2492. [PubMed] [Google Scholar]
  19. Sugino A., Higgins N. P., Brown P. O., Peebles C. L., Cozzarelli N. R. Energy coupling in DNA gyrase and the mechanism of action of novobiocin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4838–4842. doi: 10.1073/pnas.75.10.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Taylor M. J., Segall J. Characterization of factors and DNA sequences required for accurate transcription of the Saccharomyces cerevisiae 5 S RNA gene. J Biol Chem. 1985 Apr 10;260(7):4531–4540. [PubMed] [Google Scholar]
  21. Thompson R. J., Mosig G. An ATP-dependent supercoiling topoisomerase of Chlamydomonas reinhardtii affects accumulation of specific chloroplast transcripts. Nucleic Acids Res. 1985 Feb 11;13(3):873–891. doi: 10.1093/nar/13.3.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wolffe A. P., Jordan E., Brown D. D. A bacteriophage RNA polymerase transcribes through a Xenopus 5S RNA gene transcription complex without disrupting it. Cell. 1986 Feb 14;44(3):381–389. doi: 10.1016/0092-8674(86)90459-9. [DOI] [PubMed] [Google Scholar]

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