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J Biol Chem, Vol. 274, Issue 19, 13010-13016, May 7, 1999

Nuclear Ikappa Bbeta Maintains Persistent NF-kappa B Activation in HIV-1-infected Myeloid Cells

Carmela DeLucaDagger §, Louisa PetropoulosDagger parallel , Dana ZmeureanuDagger , and John HiscottDagger §parallel **

From the Dagger  Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital and the Departments of parallel  Microbiology & Immunology and § Medicine and the ** McGill AIDS Centre, McGill University, Montreal, Quebec H3T 1E2, Canada

Monocytic cells exhibit constitutive NF-kappa B activation upon infection with human immunodeficiency virus-1 (HIV-1). Because Ikappa Bbeta has been implicated in maintaining NF-kappa B·DNA binding, we sought to investigate whether Ikappa Bbeta was involved in maintaining persistent NF-kappa B activation in HIV-1-infected monocytic cell lines. Ikappa Bbeta was present in the nucleus of HIV-1-infected cells and participated in the ternary complex formation with NF-kappa B and DNA. In contrast to uninfected cells, the addition of recombinant glutathione S-transferase-Ikappa Balpha protein to preformed NF-kappa B·DNA complexes from HIV-1-infected cell extracts did not completely dissociate the complexes, suggesting that Ikappa Bbeta may protect NF-kappa B complexes from Ikappa Balpha -mediated dissociation. Immunodepletion of Ikappa Bbeta resulted in an NF-kappa B·DNA binding complex that was sensitive to Ikappa Balpha -mediated dissociation, thus demonstrating the protective role of Ikappa Bbeta . In addition, co-transfection studies with an NF-kappa B-dependent reporter construct demonstrated that Ikappa Bbeta co-expression partially alleviated inhibition of NF-kappa B-mediated gene expression by Ikappa Balpha , implying that Ikappa Bbeta can maintain transcriptionally active NF-kappa B·DNA complexes. Furthermore, constitutive phosphorylation of Ikappa Balpha was observed. Immunoprecipitation of the Ikappa B kinase (IKK) complex followed by in vitro analysis of kinase activity demonstrated that IKK was constitutively activated in HIV-1-infected myeloid cells. Thus, virus-induced constitutive IKK activation, coupled with the maintenance of a ternary NF-kappa B·DNA complex by Ikappa Bbeta , maintains persistent NF-kappa B activity in HIV-1-infected myeloid cells.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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