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Journal of Virology, January 2005, p. 95-105, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.95-105.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Induction of the RelB NF-{kappa}B Subunit by the Cytomegalovirus IE1 Protein Is Mediated via Jun Kinase and c-Jun/Fra-2 AP-1 Complexes

Xiaobo Wang and Gail E. Sonenshein*

Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts

Received 4 June 2004/ Accepted 11 August 2004

We recently demonstrated that the cytomegalovirus (CMV) immediate-early 1 (IE1) protein induces transcription of the gene encoding the RelB NF-{kappa}B subunit. The mechanism of this activation has been explored here. We report that the induction of the relB promoter by IE1 protein is mediated via activation of JNK and AP-1. The region controlling relB promoter induction was mapped to the upstream ~600-bp region between –1694 and –1096 bp. IE1 stimulated AP-1 activity in NIH 3T3 cells. Competition electrophoretic mobility shift assay (EMSA) confirmed the presence of one bona fide AP-1 element centered at –1503 bp. Introduction of a G-to-C mutation in the AP-1 binding site within the distal region of the relB promoter eliminated its activation by IE1 in both NIH 3T3 fibroblasts and vascular smooth muscle cells (SMCs). Supershift EMSA identified c-Jun, Fra-2, and c-Fos in AP-1 binding complexes in IE1 transfected NIH 3T3 cells. IE1 induced c-Jun phosphorylation, and treatment with SP600125, a selective JNK inhibitor, as well as overexpression of JNK-binding domain of JIP1, blocked IE1-mediated induction of AP-1 and relB promoter activity in NIH 3T3 cells and SMCs. Ectopic expression of c-Jun plus Fra-2, but not c-Fos, induced relB promoter activity. The relB promoter has two proximal NF-{kappa}B elements, and c-Jun/Fra-2 worked in synergy with p50/p65 NF-{kappa}B complexes. Overall, these findings demonstrate for the first time the role of AP-1 in transcriptional regulation of a gene encoding an NF-{kappa}B subunit, and its involvement in induction of RelB activity by the CMV IE1 protein.

* Corresponding author. Mailing address: Department of Biochemistry, Boston University School of Medicine, 715 Albany St., Boston, MA 02118. Phone: (617) 638-4120. Fax: (617) 638-4252. E-mail: gsonensh{at}bu.edu.

Journal of Virology, January 2005, p. 95-105, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.95-105.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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