Skip to main content
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
. 1995 Oct 10;92(21):9727–9731. doi: 10.1073/pnas.92.21.9727

DNA loops induced by cooperative binding of transcriptional activator proteins and preinitiation complexes.

J C Becker 1, A Nikroo 1, T Brabletz 1, R A Reisfeld 1
PMCID: PMC40875  PMID: 7568206

Abstract

DNA conformational changes are essential for the assembly of multiprotein complexes that contact several DNA sequence elements. An approach based on atomic force microscopy was chosen to visualize specific protein-DNA interactions occurring on eukaryotic class II nuclear gene promoters. Here we report that binding of the transcription regulatory protein Jun to linearized plasmid DNA containing the consensus AP-1 binding site upstream of a class II gene promoter leads to bending of the DNA template. This binding of Jun was found to be essential for the formation of preinitiation complexes (PICs). The cooperative binding of Jun and PIC led to looping of DNA at the protein binding sites. These loops were not seen in the absence of either PICs, Jun, or the AP-1 binding site, suggesting a direct interaction between DNA-bound Jun homodimers and proteins bound to the core promoter. This direct visualization of functional transcriptional complexes confirms the theoretical predictions for the mode of gene regulation by trans-activating proteins.

Full text

PDF
9730

Images in this article

Selected References

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

  1. Allison D. P., Bottomley L. A., Thundat T., Brown G. M., Woychik R. P., Schrick J. J., Jacobson K. B., Warmack R. J. Immobilization of DNA for scanning probe microscopy. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10129–10133. doi: 10.1073/pnas.89.21.10129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bazett-Jones D. P., Leblanc B., Herfort M., Moss T. Short-range DNA looping by the Xenopus HMG-box transcription factor, xUBF. Science. 1994 May 20;264(5162):1134–1137. doi: 10.1126/science.8178172. [DOI] [PubMed] [Google Scholar]
  3. Becker J. C., Brabletz T., Kirchner T., Conrad C. T., Bröcker E. B., Reisfeld R. A. Negative transcriptional regulation in anergic T cells. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2375–2378. doi: 10.1073/pnas.92.6.2375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buratowski S. The basics of basal transcription by RNA polymerase II. Cell. 1994 Apr 8;77(1):1–3. doi: 10.1016/0092-8674(94)90226-7. [DOI] [PubMed] [Google Scholar]
  5. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Erie D. A., Yang G., Schultz H. C., Bustamante C. DNA bending by Cro protein in specific and nonspecific complexes: implications for protein site recognition and specificity. Science. 1994 Dec 2;266(5190):1562–1566. doi: 10.1126/science.7985026. [DOI] [PubMed] [Google Scholar]
  7. Giese K., Cox J., Grosschedl R. The HMG domain of lymphoid enhancer factor 1 bends DNA and facilitates assembly of functional nucleoprotein structures. Cell. 1992 Apr 3;69(1):185–195. doi: 10.1016/0092-8674(92)90129-z. [DOI] [PubMed] [Google Scholar]
  8. Gill G., Pascal E., Tseng Z. H., Tjian R. A glutamine-rich hydrophobic patch in transcription factor Sp1 contacts the dTAFII110 component of the Drosophila TFIID complex and mediates transcriptional activation. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):192–196. doi: 10.1073/pnas.91.1.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Griffith J., Hochschild A., Ptashne M. DNA loops induced by cooperative binding of lambda repressor. Nature. 1986 Aug 21;322(6081):750–752. doi: 10.1038/322750a0. [DOI] [PubMed] [Google Scholar]
  10. Guthold M., Bezanilla M., Erie D. A., Jenkins B., Hansma H. G., Bustamante C. Following the assembly of RNA polymerase-DNA complexes in aqueous solutions with the scanning force microscope. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12927–12931. doi: 10.1073/pnas.91.26.12927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hansma H. G., Vesenka J., Siegerist C., Kelderman G., Morrett H., Sinsheimer R. L., Elings V., Bustamante C., Hansma P. K. Reproducible imaging and dissection of plasmid DNA under liquid with the atomic force microscope. Science. 1992 May 22;256(5060):1180–1184. doi: 10.1126/science.256.5060.1180. [DOI] [PubMed] [Google Scholar]
  12. Harrington R. E., Winicov I. New concepts in protein-DNA recognition: sequence-directed DNA bending and flexibility. Prog Nucleic Acid Res Mol Biol. 1994;47:195–270. doi: 10.1016/s0079-6603(08)60253-6. [DOI] [PubMed] [Google Scholar]
  13. Kahn J. D., Yun E., Crothers D. M. Detection of localized DNA flexibility. Nature. 1994 Mar 10;368(6467):163–166. doi: 10.1038/368163a0. [DOI] [PubMed] [Google Scholar]
  14. Kerppola T. K., Curran T. DNA bending by Fos and Jun: the flexible hinge model. Science. 1991 Nov 22;254(5035):1210–1214. doi: 10.1126/science.1957173. [DOI] [PubMed] [Google Scholar]
  15. Kerppola T. K., Curran T. Fos-Jun heterodimers and Jun homodimers bend DNA in opposite orientations: implications for transcription factor cooperativity. Cell. 1991 Jul 26;66(2):317–326. doi: 10.1016/0092-8674(91)90621-5. [DOI] [PubMed] [Google Scholar]
  16. Kim Y., Geiger J. H., Hahn S., Sigler P. B. Crystal structure of a yeast TBP/TATA-box complex. Nature. 1993 Oct 7;365(6446):512–520. doi: 10.1038/365512a0. [DOI] [PubMed] [Google Scholar]
  17. Klein C., Struhl K. Increased recruitment of TATA-binding protein to the promoter by transcriptional activation domains in vivo. Science. 1994 Oct 14;266(5183):280–282. doi: 10.1126/science.7939664. [DOI] [PubMed] [Google Scholar]
  18. König P., Richmond T. J. The X-ray structure of the GCN4-bZIP bound to ATF/CREB site DNA shows the complex depends on DNA flexibility. J Mol Biol. 1993 Sep 5;233(1):139–154. doi: 10.1006/jmbi.1993.1490. [DOI] [PubMed] [Google Scholar]
  19. Lin Y. S., Green M. R. Mechanism of action of an acidic transcriptional activator in vitro. Cell. 1991 Mar 8;64(5):971–981. doi: 10.1016/0092-8674(91)90321-o. [DOI] [PubMed] [Google Scholar]
  20. Lyubchenko Y., Shlyakhtenko L., Harrington R., Oden P., Lindsay S. Atomic force microscopy of long DNA: imaging in air and under water. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2137–2140. doi: 10.1073/pnas.90.6.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mills J. B., Cooper J. P., Hagerman P. J. Electrophoretic evidence that single-stranded regions of one or more nucleotides dramatically increase the flexibility of DNA. Biochemistry. 1994 Feb 22;33(7):1797–1803. doi: 10.1021/bi00173a024. [DOI] [PubMed] [Google Scholar]
  22. Patel L., Abate C., Curran T. Altered protein conformation on DNA binding by Fos and Jun. Nature. 1990 Oct 11;347(6293):572–575. doi: 10.1038/347572a0. [DOI] [PubMed] [Google Scholar]
  23. Ptashne M. Gene regulation by proteins acting nearby and at a distance. Nature. 1986 Aug 21;322(6081):697–701. doi: 10.1038/322697a0. [DOI] [PubMed] [Google Scholar]
  24. Rees W. A., Keller R. W., Vesenka J. P., Yang G., Bustamante C. Evidence of DNA bending in transcription complexes imaged by scanning force microscopy. Science. 1993 Jun 11;260(5114):1646–1649. doi: 10.1126/science.8503010. [DOI] [PubMed] [Google Scholar]
  25. Roberts S. G., Green M. R. Activator-induced conformational change in general transcription factor TFIIB. Nature. 1994 Oct 20;371(6499):717–720. doi: 10.1038/371717a0. [DOI] [PubMed] [Google Scholar]
  26. Stringer K. F., Ingles C. J., Greenblatt J. Direct and selective binding of an acidic transcriptional activation domain to the TATA-box factor TFIID. Nature. 1990 Jun 28;345(6278):783–786. doi: 10.1038/345783a0. [DOI] [PubMed] [Google Scholar]
  27. Thundat T., Allison D. P., Warmack R. J., Brown G. M., Jacobson K. B., Schrick J. J., Ferrell T. L. Atomic force microscopy of DNA on mica and chemically modified mica. Scanning Microsc. 1992 Dec;6(4):911–918. [PubMed] [Google Scholar]
  28. Tjian R., Maniatis T. Transcriptional activation: a complex puzzle with few easy pieces. Cell. 1994 Apr 8;77(1):5–8. doi: 10.1016/0092-8674(94)90227-5. [DOI] [PubMed] [Google Scholar]
  29. Travers A., Klug A. Nucleoprotein complexes. DNA wrapping and writhing. 1987 May 28-Jun 3Nature. 327(6120):280–281. doi: 10.1038/327280a0. [DOI] [PubMed] [Google Scholar]
  30. Wang J. C., Giaever G. N. Action at a distance along a DNA. Science. 1988 Apr 15;240(4850):300–304. doi: 10.1126/science.3281259. [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