The Ets Transcription Factors Interact with Each Other and with the c-Fos/c-Jun Complex via Distinct Protein Domains in a DNA-dependent and -independent Manner*
Abstract
The transcription factors Fos, Jun, and Ets regulate the expression of human stromelysin-1 and collagenase-1 genes. Recently, we found that ERG, an Ets family member, activates collagenase-1 gene but not stromelysin-1 by physically interacting with c-Fos/c-Jun. Interestingly, ERG binds to stromelysin-1 promoter and represses its activation by ETS2. Here, to investigate the molecular mechanism of this regulation, we have used an in vitroprotein-protein interaction assay and studied the transcription factor interactions of ETS2. We found that ETS2 could weakly associate within vitro synthesized ETS1, c-Fos, and c-Jun and strongly with c-Fos/c-Jun complex and ERG via several distinct ETS2 domains including the C-terminal region that contains the DNA-binding domain. Strikingly, these interactions were stabilized in vitro by DNA as they were inhibited by ethidium bromide. Both the N-terminal region, comprising the transactivation domain, and the C-terminal region of ETS2 associated with ERG and, interestingly, the interaction of ERG through the transactivation domain of ETS2 was DNA-independent. The DNA-dependent interaction of ETS2 with c-Fos/c-Jun was enhanced by specific DNA fragments requiring two Ets-binding sites of the stromelysin-1 promoter. Using the two hybrid system, we also demonstrated that ETS2 interacts with c-Jun or ERG in vivo.
Footnotes
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↵* This work was supported by Grant ARC 1093 (to G. B.) from the Association pour la Recherche sur le Cancer (ARC) and by the Institut Pasteur of Lille.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵‡ Recipient of a scholarship from the ARC.
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↵§ To whom correspondence should be addressed. Tel.: 33 3 20 87 10 97; Fax: 33 3 20 87 11 11; E-mail: gbuttice @ infobiogen.fr.
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↵1 The abbreviations used are: DBD, DNA-binding domain; TA, transactivation; GST, glutathione S-transferase; PAGE, polyacrylamide gel electrophoresis; EtBr, ethidium bromide; ID, inhibitory domain(s); EBS, ETS-binding sites; PCR, polymerase chain reaction.
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↵2 J. P. Basuyaux and G. Butticè, unpublished data.
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- Received June 16, 1997.
