Skip to main content
PMC 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
. 1993 Apr 15;90(8):3339–3342. doi: 10.1073/pnas.90.8.3339

Z-DNA binding protein from chicken blood nuclei.

A G Herbert 1, J R Spitzner 1, K Lowenhaupt 1, A Rich 1
PMCID: PMC46295  PMID: 8475077

Abstract

A protein (Z alpha) that appears to be highly specific for the left-handed Z-DNA conformer has been identified in chicken blood nuclear extracts. Z alpha activity is measured in a band-shift assay by using a radioactive probe consisting of a (dC-dG)35 oligomer that has 50% of the deoxycytosines replaced with 5-bromodeoxycytosine. In the presence of 10 mM Mg2+, the probe converts to the Z-DNA conformation and is bound by Z alpha. The binding of Z alpha to the radioactive probe is specifically blocked by competition with linear poly(dC-dG) stabilized in the Z-DNA form by chemical bromination but not by B-form poly(dC-dG) or boiled salmon-sperm DNA. In addition, the binding activity of Z alpha is competitively blocked by supercoiled plasmids containing a Z-DNA insert but not by either the linearized plasmid or by an equivalent amount of the parental supercoiled plasmid without the Z-DNA-forming insert. Z alpha can be crosslinked to the 32P-labeled brominated probe with UV light, allowing us to estimate that the minimal molecular mass of Z alpha is 39 kDa.

Full text

PDF
3342

Images in this article

3340Fig. 1
on p.3340
3340Fig. 2
on p.3340
3341Fig. 3
on p.3341
3341Fig. 4
on p.3341

Selected References

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

  1. Behe M., Felsenfeld G. Effects of methylation on a synthetic polynucleotide: the B--Z transition in poly(dG-m5dC).poly(dG-m5dC). Proc Natl Acad Sci U S A. 1981 Mar;78(3):1619–1623. doi: 10.1073/pnas.78.3.1619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blaho J. A., Wells R. D. Left-handed Z-DNA binding by the recA protein of Escherichia coli. J Biol Chem. 1987 May 5;262(13):6082–6088. [PubMed] [Google Scholar]
  3. Gilmour D. S., Thomas G. H., Elgin S. C. Drosophila nuclear proteins bind to regions of alternating C and T residues in gene promoters. Science. 1989 Sep 29;245(4925):1487–1490. doi: 10.1126/science.2781290. [DOI] [PubMed] [Google Scholar]
  4. Gut S. H., Bischoff M., Hobi R., Kuenzle C. C. Z-DNA-binding proteins from bull testis. Nucleic Acids Res. 1987 Dec 10;15(23):9691–9705. doi: 10.1093/nar/15.23.9691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Haniford D. B., Pulleyblank D. E. The in-vivo occurrence of Z DNA. J Biomol Struct Dyn. 1983 Dec;1(3):593–609. doi: 10.1080/07391102.1983.10507467. [DOI] [PubMed] [Google Scholar]
  6. Kim J. I., Heuser J., Cox M. M. Enhanced recA protein binding to Z DNA represents a kinetic perturbation of a general duplex DNA binding pathway. J Biol Chem. 1989 Dec 25;264(36):21848–21856. [PubMed] [Google Scholar]
  7. Krishna P., Kennedy B. P., Waisman D. M., van de Sande J. H., McGhee J. D. Are many Z-DNA binding proteins actually phospholipid-binding proteins? Proc Natl Acad Sci U S A. 1990 Feb;87(4):1292–1295. doi: 10.1073/pnas.87.4.1292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kłysik J., Stirdivant S. M., Larson J. E., Hart P. A., Wells R. D. Left-handed DNA in restriction fragments and a recombinant plasmid. Nature. 1981 Apr 23;290(5808):672–677. doi: 10.1038/290672a0. [DOI] [PubMed] [Google Scholar]
  9. Lafer E. M., Sousa R. J., Rich A. Z-DNA-binding proteins in Escherichia coli purification, generation of monoclonal antibodies and gene isolation. J Mol Biol. 1988 Sep 20;203(2):511–516. doi: 10.1016/0022-2836(88)90017-4. [DOI] [PubMed] [Google Scholar]
  10. Muller M. T., Spitzner J. R., DiDonato J. A., Mehta V. B., Tsutsui K., Tsutsui K. Single-strand DNA cleavages by eukaryotic topoisomerase II. Biochemistry. 1988 Nov 1;27(22):8369–8379. doi: 10.1021/bi00422a012. [DOI] [PubMed] [Google Scholar]
  11. Möller A., Nordheim A., Kozlowski S. A., Patel D. J., Rich A. Bromination stabilizes poly(dG-dC) in the Z-DNA form under low-salt conditions. Biochemistry. 1984 Jan 3;23(1):54–62. doi: 10.1021/bi00296a009. [DOI] [PubMed] [Google Scholar]
  12. Peck L. J., Nordheim A., Rich A., Wang J. C. Flipping of cloned d(pCpG)n.d(pCpG)n DNA sequences from right- to left-handed helical structure by salt, Co(III), or negative supercoiling. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4560–4564. doi: 10.1073/pnas.79.15.4560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pohl F. M., Jovin T. M. Salt-induced co-operative conformational change of a synthetic DNA: equilibrium and kinetic studies with poly (dG-dC). J Mol Biol. 1972 Jun 28;67(3):375–396. doi: 10.1016/0022-2836(72)90457-3. [DOI] [PubMed] [Google Scholar]
  14. Rahmouni A. R., Wells R. D. Stabilization of Z DNA in vivo by localized supercoiling. Science. 1989 Oct 20;246(4928):358–363. doi: 10.1126/science.2678475. [DOI] [PubMed] [Google Scholar]
  15. Rich A., Nordheim A., Wang A. H. The chemistry and biology of left-handed Z-DNA. Annu Rev Biochem. 1984;53:791–846. doi: 10.1146/annurev.bi.53.070184.004043. [DOI] [PubMed] [Google Scholar]
  16. Rohner K. J., Hobi R., Kuenzle C. C. Z-DNA-binding proteins. Identification critically depends on the proper choice of ligands. J Biol Chem. 1990 Nov 5;265(31):19112–19115. [PubMed] [Google Scholar]
  17. Schroth G. P., Chou P. J., Ho P. S. Mapping Z-DNA in the human genome. Computer-aided mapping reveals a nonrandom distribution of potential Z-DNA-forming sequences in human genes. J Biol Chem. 1992 Jun 15;267(17):11846–11855. [PubMed] [Google Scholar]
  18. Sheardy R. D. Preliminary spectroscopic characterization of a synthetic DNA oligomer containing a B-Z junction at high salt. Nucleic Acids Res. 1988 Feb 11;16(3):1153–1167. doi: 10.1093/nar/16.3.1153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tripathi J., Brahmachari S. K. Distribution of simple repetitive (TG/CA)n and (CT/AG)n sequences in human and rodent genomes. J Biomol Struct Dyn. 1991 Oct;9(2):387–397. doi: 10.1080/07391102.1991.10507919. [DOI] [PubMed] [Google Scholar]
  20. Wang A. H., Quigley G. J., Kolpak F. J., Crawford J. L., van Boom J. H., van der Marel G., Rich A. Molecular structure of a left-handed double helical DNA fragment at atomic resolution. Nature. 1979 Dec 13;282(5740):680–686. doi: 10.1038/282680a0. [DOI] [PubMed] [Google Scholar]
  21. Wittig B., Wölfl S., Dorbic T., Vahrson W., Rich A. Transcription of human c-myc in permeabilized nuclei is associated with formation of Z-DNA in three discrete regions of the gene. EMBO J. 1992 Dec;11(12):4653–4663. doi: 10.1002/j.1460-2075.1992.tb05567.x. [DOI] [PMC free article] [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