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. 1984 Oct 25;12(20):7753–7769. doi: 10.1093/nar/12.20.7753

Competition between Xenopus satellite I sequences and Pol III genes for stable transcription complex formation.

D L Andrews, L Millstein, B A Hamkalo, J M Gottesfeld
PMCID: PMC320198  PMID: 6093052

Abstract

We have constructed hybrid plasmids bearing both Xenopus 5S RNA genes and satellite I sequences in order to test the effect of satellite DNA on 5S gene transcription. Satellite sequences inactivate 5S transcription in both HeLa S100 and Xenopus oocyte microinjection transcription assays. Inactivation of 5S transcription by satellite DNA is observed both in cis and in trans. Transcription of a tRNA gene is also precluded by satellite I DNA. The Xenopus satellite I repeat contains an RNA polymerase III transcription unit which is highly active in both assay systems. This promoter element is 10- to 25-fold more efficient than the 5S gene in transcription competition assays. This quantitative difference in affinity for transcription components may explain the inactivation of 5S transcription by satellite sequences. The satellite I promoter forms stable transcription complexes in vitro which do not dissociate for at least 30 rounds of transcription. Although stable complex formation on the satellite promoter is largely temperature independent over the range of 0-20 degrees, complex formation on both 5S and tRNA genes exhibits a steep temperature dependence characteristic of DNA helix unwinding. The DNA sequence requirements for stable complex formation on 5S genes have been determined using 5' deletion mutants.

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Selected References

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

  1. Ackerman E. J. Molecular cloning and sequencing of OAX DNA: an abundant gene family transcribed and activated in Xenopus oocytes. EMBO J. 1983;2(8):1417–1422. doi: 10.1002/j.1460-2075.1983.tb01600.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bogenhagen D. F., Brown D. D. Nucleotide sequences in Xenopus 5S DNA required for transcription termination. Cell. 1981 Apr;24(1):261–270. doi: 10.1016/0092-8674(81)90522-5. [DOI] [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. Brutlag D. L. Molecular arrangement and evolution of heterochromatic DNA. Annu Rev Genet. 1980;14:121–144. doi: 10.1146/annurev.ge.14.120180.001005. [DOI] [PubMed] [Google Scholar]
  5. Ciliberto G., Traboni C., Cortese R. Relationship between the two components of the split promoter of eukaryotic tRNA genes. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1921–1925. doi: 10.1073/pnas.79.6.1921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Diaz M. O., Barsacchi-Pilone G., Mahon K. A., Gall J. G. Transcripts from both strands of a satellite DNA occur on lampbrush chromosome loops of the newt Notophthalmus. Cell. 1981 Jun;24(3):649–659. doi: 10.1016/0092-8674(81)90091-x. [DOI] [PubMed] [Google Scholar]
  7. Dingermann T., Sharp S., Schaack J., Söll D. Stable transcription complex formation of eukaryotic tRNA genes is dependent on a limited separation of the two intragenic control regions. J Biol Chem. 1983 Sep 10;258(17):10395–10402. [PubMed] [Google Scholar]
  8. Galli G., Hofstetter H., Birnstiel M. L. Two conserved sequence blocks within eukaryotic tRNA genes are major promoter elements. Nature. 1981 Dec 17;294(5842):626–631. doi: 10.1038/294626a0. [DOI] [PubMed] [Google Scholar]
  9. Gamper H. B., Hearst J. E. Size of the unwound region of DNA in Escherichia coli RNA polymerase and calf thymus RNA polymerase II ternary complexes. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):455–461. doi: 10.1101/sqb.1983.047.01.054. [DOI] [PubMed] [Google Scholar]
  10. Jamrich M., Warrior R., Steele R., Gall J. G. Transcription of repetitive sequences on Xenopus lampbrush chromosomes. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3364–3367. doi: 10.1073/pnas.80.11.3364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lam B. S., Carroll D. Tandemly repeated DNA sequences from Xenopus laevis. I. Studies on sequence organization and variation in satellite 1 DNA (741 base-pair repeat). J Mol Biol. 1983 Apr 25;165(4):567–585. doi: 10.1016/s0022-2836(83)80267-8. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Sakonju S., Brown D. D. Contact points between a positive transcription factor and the Xenopus 5S RNA gene. Cell. 1982 Dec;31(2 Pt 1):395–405. doi: 10.1016/0092-8674(82)90133-7. [DOI] [PubMed] [Google Scholar]
  15. Sakonju S., Brown D. D., Engelke D., Ng S. Y., Shastry B. S., Roeder R. G. The binding of a transcription factor to deletion mutants of a 5S ribosomal RNA gene. Cell. 1981 Mar;23(3):665–669. doi: 10.1016/0092-8674(81)90429-3. [DOI] [PubMed] [Google Scholar]
  16. Travers A. RNA polymerase--promoter interactions: some general principles. Cell. 1974 Oct;3(2):97–104. doi: 10.1016/0092-8674(74)90112-3. [DOI] [PubMed] [Google Scholar]
  17. Wakefield L., Ackerman E., Gurdon J. B. The activation of RNA synthesis by somatic nuclei injected into amphibian oocytes. Dev Biol. 1983 Feb;95(2):468–475. doi: 10.1016/0012-1606(83)90048-9. [DOI] [PubMed] [Google Scholar]
  18. Weil P. A., Segall J., Harris B., Ng S. Y., Roeder R. G. Faithful transcription of eukaryotic genes by RNA polymerase III in systems reconstituted with purified DNA templates. J Biol Chem. 1979 Jul 10;254(13):6163–6173. [PubMed] [Google Scholar]
  19. Wormington W. M., Bogenhagen D. F., Jordan E., Brown D. D. A quantitative assay for Xenopus 5S RNA gene transcription in vitro. Cell. 1981 Jun;24(3):809–817. doi: 10.1016/0092-8674(81)90106-9. [DOI] [PubMed] [Google Scholar]

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