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Molecular and Cellular Biology, February 1999, p. 959-966, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Essential Role of Interferon Regulatory Factor 3 in Direct Activation of RANTES Chemokine Transcription

Rongtuan Lin,1,2,* Christophe Heylbroeck,1,3 Pierre Genin,1,3 Paula M. Pitha,4 and John Hiscott1,2,3

Terry Fox Molecular Oncology Group, Lady Davis Institute for Medical Research,1 and Departments of Microbiology and Immunology3 and Medicine,2 McGill University, Montreal, Canada H3T 1E2, and Oncology Center, School of Medicine, Johns Hopkins University, Baltimore, Maryland 212314

Received 1 July 1998/Returned for modification 10 September 1998/Accepted 27 October 1998

Localized and systemic cytokine production in virus-infected cells play an important role in the outcome of viral infection and pathogenicity. Activation of the interferon regulatory factors (IRF) in turn is a critical mediator of cytokine gene transcription. Recent studies have focused on the 55-kDa IRF-3 gene product as a direct transcriptional regulator of type 1 interferon (IFN-alpha and IFN-beta ) activation in response to virus infection. Virus infection induces phosphorylation of IRF-3 on specific C-terminal serine residues and permits cytoplasmic-to-nuclear translocation of IRF-3, activation of DNA binding and transactivation potential, and association with the CBP/p300 coactivator. We previously generated constitutively active [IRF-3(5D)] and dominant-negative forms of IRF-3 that control IFN-beta and IFN-alpha gene expression. In an effort to characterize the range of immunoregulatory genes controlled by IRF-3, we now demonstrate that endogenous human RANTES gene transcription is directly induced in tetracycline-inducible IRF-3(5D)-expressing cells or paramyxovirus-infected cells. We also show that a dominant-negative IRF-3 mutant inhibits virus-induced expression of the RANTES promoter. Specific mutagenesis of overlapping ISRE-like sites located between nucleotides -123 and -96 in the RANTES promoter reduces virus-induced and IRF-3-dependent activation. These studies broaden the range of IRF-3 immunoregulatory target genes to include at least one member of the chemokine superfamily.

* Corresponding author. Mailing address: Lady Davis Institute for Medical Research, 3755 Cote Ste. Catherine, Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8222, ext. 4509. Fax: (514) 340-7576. E-mail: mdli{at}musica.mcgill.ca.

Molecular and Cellular Biology, February 1999, p. 959-966, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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