Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities


One of the biggest challenges facing gene therapy is the development of vectors that direct the activity of therapeutic genes specifically to the sites of disease. To achieve this goal, the restriction of transgene transcription via synthetic promoters that are endowed with multiple specificities represents a particularly promising strategy. Towards this end, we have developed a generally applicable strategy (DCTF system) where a synthetic promoter is driven by an artificial heterodimeric transcription factor whose DNA-binding and transactivating subunits are expressed from two promoters with different selectivity. A crucial determinant of the DCTF system is the heterodimerization interface that should provide for a high affinity interaction without interference by endogenous proteins. Here, we describe such a dimerization system based on engineered Fos and Jun leucine zippers. We show the usefulness of this system for the combination of cell type-specific and cell cycle-regulated transcription and demonstrate its functionality in an in vivo setting.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3


  1. Jérôme V, Müller R . Tissue-specific, cell cycle-regulated chimeric transcription factors for the targeting of gene expression to tumor cells Hum Gene Ther 1998 9: 2653–2659

    Article  Google Scholar 

  2. Busch SJ, Sassone-Corsi P . Dimers, leucine zippers and DNA-binding domains Trends Genet 1990 6: 36–40

    CAS  Article  Google Scholar 

  3. Neuberg M, Adamkiewicz J, Hunter JB, Müller R . A Fos protein containing the Jun leucine zipper forms a homodimer which binds to the AP1 binding site Nature 1989 341: 243–245

    CAS  Article  Google Scholar 

  4. Kouzarides T, Ziff E . Leucine zippers of fos, jun and GCN4 dictate dimerization specificity and thereby control DNA binding Nature 1989 340: 568–571

    CAS  Article  Google Scholar 

  5. Schmidt-Dörr T et al. Construction, purification and characterization of a hybrid protein comprising the DNA binding domain of the LexA repressor and the Jun leucine zipper: a circular dichroism and mutagenesis study Biochemistry 1991 30: 9657–9664

    Article  Google Scholar 

  6. Kouzarides T, Ziff E . The role of the leucine zipper in the fos–jun interaction Nature 1988 336: 646–651

    CAS  Article  Google Scholar 

  7. O'Shea EK, Rutkowski R, Stafford WFD, Kim PS . Preferential heterodimer formation by isolated leucine zippers from fos and jun Science 1989 245: 646–648

    CAS  Article  Google Scholar 

  8. Schuermann M et al. The leucine repeat motif in Fos protein mediates complex formation with Jun/AP-1 and is required for transformation Cell 1989 56: 507–516

    CAS  Article  Google Scholar 

  9. Oas TG et al. Secondary structure of a leucine zipper determined by nuclear magnetic resonance spectroscopy Biochemistry 1990 29: 2891–2894

    CAS  Article  Google Scholar 

  10. Schuermann M, Hunter JB, Hennig G, Müller R . Non-leucine residues in the leucine repeats of Fos and Jun contribute to the stability and determine the specifity of dimerization Nucleic Acids Res 1991 19: 739–746

    CAS  Article  Google Scholar 

  11. O'Shea EK, Rutkowski R, Kim PS . Mechanism of specificity in the Fos-Jun oncoprotein heterodimer Cell 1992 68: 699–708

    CAS  Article  Google Scholar 

  12. Glover JN, Harrison SC . Crystal structure of the heterodimeric bZIP transcription factor c-Fos-c-Jun bound to DNA Nature 1995 373: 257–261

    CAS  Article  Google Scholar 

  13. Triezenberg SJ, Kingsbury RC, McKnight SL . Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression Genes Dev 1988 2: 718–729

    CAS  Article  Google Scholar 

  14. Chasman DI, Kornberg RD . GAL4 protein: purification, asssociation with GAL80 protein, and conserved domain structure Mol Cell Biol 1990 10: 2916–2923

    CAS  Article  Google Scholar 

  15. Dang CV et al. Intracellular leucine zipper interactions suggest c-Myc hetero-oligomerization Mol Cell Biol 1991 11: 954–962

    CAS  Article  Google Scholar 

  16. Davidson JN et al. The evolutionary history of the first three enzymes in pyrimidine biosynthesis BioEssays 1993 15: 157–164

    CAS  Article  Google Scholar 

  17. Gontero B et al. Structural and functional properties of a multi-enzyme complex from spinach chloroplasts. 2. Modulation of the kinetic properties of enzymes in the aggregated state Eur J Biochem 1993 217: 1075–1082

    CAS  Article  Google Scholar 

  18. Walczak H et al. Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo Nature Med 1999 5: 157–163

    CAS  Article  Google Scholar 

  19. Kalderon D, Roberts BL, Richardson WD, Smith AE . A short amino acid sequence able to specify nuclear location Cell 1984 39: 499–509

    CAS  Article  Google Scholar 

  20. Ransone LJ, Verma IM . Nuclear proto-oncogenes fos and jun Annu Rev Cell Biol 1990 6: 539–557

    CAS  Article  Google Scholar 

  21. Bean MA et al. Cell-mediated cytotoxicity for bladder carcinoma: evaluation of a workshop Cancer Res 1975 35: 2902–2913

    CAS  PubMed  Google Scholar 

  22. Ausubel I, Frederick M . Current Protocols in Molecular Biology John Wiley & Sons: New York 1991

  23. Cornil I, Man S, Fernandez B, Kerbel RS . Enhanced tumorigenicity, melanogenesis, and metastases of a human malignant melanoma after subdermal implantation in nude mice J Natl Cancer Inst 1989 81: 938–944

    CAS  Article  Google Scholar 

  24. Coll JL et al. Antitumor activity of bax and p53 naked gene transfer in lung cancer: in vitro and in vivo analysis Hum Gene Ther 1998 9: 2063–2074

    CAS  Article  Google Scholar 

Download references


We are grateful to I Hart for the MeWo cell line, to N Favrot for the H322 cell line and to M Krause for oligonucleotide synthesis. We thank Simone Schmitt for performing the in vitro translation association assay. This work was supported by the Dr Mildred Scheel Stiftung.

Author information

Authors and Affiliations


Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jérôme, V., Müller, R. A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities. Gene Ther 8, 725–729 (2001).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • cell cycle regulation
  • DCTF system
  • leucine zipper
  • tissue-specific promoter
  • transcriptional targeting

Further reading


Quick links