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. 1990 Feb 25;18(4):1021–1029. doi: 10.1093/nar/18.4.1021

Reaction parameters of TFIIIA-induced supercoiling catalyzed by a Xenopus laevis cell-free extract.

J A Sekiguchi 1, E B Kmiec 1
PMCID: PMC330359  PMID: 2315014

Abstract

In addition to its fundamental role of nucleating the formation of stable transcription complexes, the Xenopus laevis 5S RNA specific transcription factor, TFIIIA, promotes a variety of DNA-associated metabolic reactions. We report that TFIIIA can induce a DNA supercoiling catalyzed by the Xenopus laevis S-150 cell-free extract on plasmids containing a single copy of the Xenopus 5S RNA gene (somatic-type). Stimulated supercoiling occurs in the presence of high concentrations of ATP (4 mM) and at a factor to DNA ratio of 1 through a mechanism most likely involving type I topoisomerase. The highest level of stimulated supercoiling occurs when TFIIIA is incubated with DNA prior to the addition of the S-150 extract. Taken together, the experiments outlined in this report establish a reliable and seminal system in which TFIIIA-induced DNA supercoiling can be observed reproducibly.

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Selected References

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

  1. Almouzni G., Méchali M. Assembly of spaced chromatin promoted by DNA synthesis in extracts from Xenopus eggs. EMBO J. 1988 Mar;7(3):665–672. doi: 10.1002/j.1460-2075.1988.tb02861.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersen J., Delihas N. Characterization of RNA-protein interactions in 7 S ribonucleoprotein particles from Xenopus laevis oocytes. J Biol Chem. 1986 Feb 25;261(6):2912–2917. [PubMed] [Google Scholar]
  3. Bieker J. J., Roeder R. G. Physical properties and DNA-binding stoichiometry of a 5 S gene-specific transcription factor. J Biol Chem. 1984 May 25;259(10):6158–6164. [PubMed] [Google Scholar]
  4. Birkenmeier E. H., Brown D. D., Jordan E. A nuclear extract of Xenopus laevis oocytes that accurately transcribes 5S RNA genes. Cell. 1978 Nov;15(3):1077–1086. doi: 10.1016/0092-8674(78)90291-x. [DOI] [PubMed] [Google Scholar]
  5. Dignam J. D., Martin P. L., Shastry B. S., Roeder R. G. Eukaryotic gene transcription with purified components. Methods Enzymol. 1983;101:582–598. doi: 10.1016/0076-6879(83)01039-3. [DOI] [PubMed] [Google Scholar]
  6. Engelke D. R., Ng S. Y., Shastry B. S., Roeder R. G. Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes. Cell. 1980 Mar;19(3):717–728. doi: 10.1016/s0092-8674(80)80048-1. [DOI] [PubMed] [Google Scholar]
  7. Glikin G. C., Ruberti I., Worcel A. Chromatin assembly in Xenopus oocytes: in vitro studies. Cell. 1984 May;37(1):33–41. doi: 10.1016/0092-8674(84)90298-8. [DOI] [PubMed] [Google Scholar]
  8. Gottesfeld J., Bloomer L. S. Assembly of transcriptionally active 5S RNA gene chromatin in vitro. Cell. 1982 Apr;28(4):781–791. doi: 10.1016/0092-8674(82)90057-5. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Kmiec E. B., Sekiguchi J. M., Cole A. D. Studies on the ATP requirements of in vitro chromatin assembly. Biochem Cell Biol. 1989 Aug;67(8):443–454. doi: 10.1139/o89-070. [DOI] [PubMed] [Google Scholar]
  12. Kmiec E. B. Transcription factor IIIA and 5S DNA supercoiling in vitro: toward a purified system. Cell. 1988 Sep 23;54(7):919–920. doi: 10.1016/0092-8674(88)90104-3. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Knezetic J. A., Luse D. S. The presence of nucleosomes on a DNA template prevents initiation by RNA polymerase II in vitro. Cell. 1986 Apr 11;45(1):95–104. doi: 10.1016/0092-8674(86)90541-6. [DOI] [PubMed] [Google Scholar]
  15. Lassar A. B., Martin P. L., Roeder R. G. Transcription of class III genes: formation of preinitiation complexes. Science. 1983 Nov 18;222(4625):740–748. doi: 10.1126/science.6356356. [DOI] [PubMed] [Google Scholar]
  16. Liu L. F., Wang J. C. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. doi: 10.1073/pnas.84.20.7024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller J., McLachlan A. D., Klug A. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J. 1985 Jun;4(6):1609–1614. doi: 10.1002/j.1460-2075.1985.tb03825.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ng S. Y., Parker C. S., Roeder R. G. Transcription of cloned Xenopus 5S RNA genes by X. laevis RNA polymerase III in reconstituted systems. Proc Natl Acad Sci U S A. 1979 Jan;76(1):136–140. doi: 10.1073/pnas.76.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pelham H. R., Wormington W. M., Brown D. D. Related 5S RNA transcription factors in Xenopus oocytes and somatic cells. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1760–1764. doi: 10.1073/pnas.78.3.1760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Peterson R. C., Doering J. L., Brown D. D. Characterization of two xenopus somatic 5S DNAs and one minor oocyte-specific 5S DNA. Cell. 1980 May;20(1):131–141. doi: 10.1016/0092-8674(80)90241-x. [DOI] [PubMed] [Google Scholar]
  21. Razvi F., Gargiulo G., Worcel A. A simple procedure for parallel sequence analysis of both strands of 5'-labeled DNA. Gene. 1983 Aug;23(2):175–183. doi: 10.1016/0378-1119(83)90049-5. [DOI] [PubMed] [Google Scholar]
  22. Reeves R. Transcriptionally active chromatin. Biochim Biophys Acta. 1984 Sep 10;782(4):343–393. doi: 10.1016/0167-4781(84)90044-7. [DOI] [PubMed] [Google Scholar]
  23. Reynolds W. F., Gottesfeld J. M. 5S rRNA gene transcription factor IIIA alters the helical configuration of DNA. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1862–1866. doi: 10.1073/pnas.80.7.1862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ruberti I., Worcel A. Mechanism of chromatin assembly in Xenopus oocytes. J Mol Biol. 1986 Jun 5;189(3):457–476. doi: 10.1016/0022-2836(86)90317-7. [DOI] [PubMed] [Google Scholar]
  25. Schlissel M. S., Brown D. D. The transcriptional regulation of Xenopus 5s RNA genes in chromatin: the roles of active stable transcription complexes and histone H1. Cell. 1984 Jul;37(3):903–913. doi: 10.1016/0092-8674(84)90425-2. [DOI] [PubMed] [Google Scholar]
  26. Sekiguchi J. M., Swank R. A., Kmiec E. B. Changes in DNA topology can modulate in vitro transcription of certain RNA polymerase III genes. Mol Cell Biochem. 1989 Feb 21;85(2):123–133. doi: 10.1007/BF00577108. [DOI] [PubMed] [Google Scholar]
  27. Shimamura A., Tremethick D., Worcel A. Characterization of the repressed 5S DNA minichromosomes assembled in vitro with a high-speed supernatant of Xenopus laevis oocytes. Mol Cell Biol. 1988 Oct;8(10):4257–4269. doi: 10.1128/mcb.8.10.4257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sinden R. R., Carlson J. O., Pettijohn D. E. Torsional tension in the DNA double helix measured with trimethylpsoralen in living E. coli cells: analogous measurements in insect and human cells. Cell. 1980 Oct;21(3):773–783. doi: 10.1016/0092-8674(80)90440-7. [DOI] [PubMed] [Google Scholar]
  29. Smith D. R., Jackson I. J., Brown D. D. Domains of the positive transcription factor specific for the Xenopus 5S RNA gene. Cell. 1984 Jun;37(2):645–652. doi: 10.1016/0092-8674(84)90396-9. [DOI] [PubMed] [Google Scholar]
  30. Wolffe A. P., Andrews M. T., Crawford E., Losa R., Brown D. D. Negative supercoiling is not required for 5S RNA transcription in vitro. Cell. 1987 May 8;49(3):301–303. doi: 10.1016/0092-8674(87)90279-0. [DOI] [PubMed] [Google Scholar]
  31. Worcel A. Transcription factor IIIA and 5S DNA supercoiling in vitro. Cell. 1988 Sep 23;54(7):919–920. doi: 10.1016/0092-8674(88)90103-1. [DOI] [PubMed] [Google Scholar]
  32. Workman J. L., Roeder R. G. Binding of transcription factor TFIID to the major late promoter during in vitro nucleosome assembly potentiates subsequent initiation by RNA polymerase II. Cell. 1987 Nov 20;51(4):613–622. doi: 10.1016/0092-8674(87)90130-9. [DOI] [PubMed] [Google Scholar]
  33. Xing Y. Y., Worcel A. The C-terminal domain of transcription factor IIIA interacts differently with different 5S RNA genes. Mol Cell Biol. 1989 Feb;9(2):499–514. doi: 10.1128/mcb.9.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]

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