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Molecular and Cellular Biology, July 2002, p. 4952-4964, Vol. 22, No. 13
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.13.4952-4964.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Promoter Specificity and Biological Activity of Tethered AP-1 Dimers

Latifa Bakiri,1,{dagger} Koichi Matsuo,2,{ddagger} Marta Wisniewska,1,3 Erwin F. Wagner,2 and Moshe Yaniv1*

Unité Expression Génétique et Maladies, CNRS URA 1644, Institut Pasteur, 75724 Paris Cedex 15, France,1 Research Institute of Molecular Pathology (IMP), 1030 Vienna, Austria,2 Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland3

Received 7 March 2002/ Accepted 3 April 2002

Activator protein 1 (AP-1) is a group of dimeric transcription factors composed of Jun, Fos, and ATF family proteins. Both gain- and loss-of-function studies have revealed specific roles for individual AP-1 components in cell proliferation, differentiation, apoptosis, and other biological processes. However, little is known about the functions of specific AP-1 dimers. To test the importance of AP-1 composition in transcriptional activation, AP-1 monomers were joined via a flexible polypeptide tether to force specific pairing. The resultant single-chain AP-1 molecules showed DNA binding specificity and transcriptional activation potentials similar to those of untethered dimers, even in the presence of dominant-negative AP-1 monomers. c-Jun-containing dimers showed distinct promoter specificity in transient-transfection experiments, depending on the Fos, Fra, or ATF partner. When stably expressed in NIH 3T3 cells, c-Jun~Fra2, but not c-Jun~Fra1 and c-Jun~cFos (the tilde indicates a tethered dimer), inhibited G0 arrest at confluency and under low-serum conditions and specifically activated cyclin A expression. These data suggest that the choice of dimerization partner defines the role of c-Jun in gene activation and cell cycle regulation and that single-chain AP-1 molecules provide a powerful tool for assessing the role of specific AP-1 dimers.

* Corresponding author. Mailing address: Unité Expression génétique et Maladies, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France. Phone: 00 33 1 45 68 85 12. Fax: 00 33 1 40 61 30 33. E-mail: yaniv{at}pasteur.fr.

{dagger} Present address: Research Institute of Molecular Pathology (IMP), 1030 Vienna, Austria.

{ddagger} Present address: Department of Geriatric Research, National Institute for Longevity Sciences, Morioka, Obu, Aichi 474-8522, Japan.

Molecular and Cellular Biology, July 2002, p. 4952-4964, Vol. 22, No. 13
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.13.4952-4964.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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