Mechanism of inactivation of NF-κB by a viral homologue of IκBα. Signal-induced release of IκBα results in binding of the viral homologue to NF-κB

Stephen W. G. Tait, Elizabeth B. Reid, David R. Greaves, Thomas E. Wileman, Penelope P. Powell

Research output: Contribution to journalArticlepeer-review

75 Citations (Scopus)

Abstract

Activation of the nuclear factor κB plays a key role in viral pathogenesis, resulting in inflammation and modulation of the immune response. We have previously shown that A238L, an open reading frame from African swine fever virus (ASFV), encoding a protein with 46% homology to porcine IκBα exerts a potent anti-inflammatory effect in host macrophages, where it down-regulates NF-κB-dependent gene transcription and proinflammatory cytokine production. This paper reveals the mechanism of suppression of NF-κB activity by A238Lp. A238Lp is synthesized throughout infection as two molecular mass forms of 28 and 32 kDa, and vaccinia-mediated expression of A238L demonstrated that both proreins are produced from a single gene. Significantly, the higher 32-kDa form of A238L, but not the 28-kDa form, interacts directly with Re1A, the 65-kDa subunit of NF-κβ, indicating that the binding is dependent on a posttranslational modification. Immunoprecipitation analysis shows the NF-κB p65-A238L p32 heterodimer is a separate complex from NF-κB-IκBα, and it resides in the cytoplasm. Moreover, we show that ASFV infection stimulates the NFκB signal transduction pathway, which results in the rapid degradation of endogenous IκBα, although both forms of A238Lp are resistant to stimulus-induced degradation. Using the proteasome inhibitor MG132, we show that when degradation of IκBα is inhibited, A238Lp binding to NF-κB p65 is reduced. The results suggest that the virus exploits its activation of the NF-κB pathway to enable its own IκB homologue to bind to NF-κB p65. Last, we show that synthesis of IκBα is increased during ASFV infection, indicating Re1A-independent transcription of the IκBα gene.

Original languageEnglish
Pages (from-to)34656-34664
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number44
DOIs
Publication statusPublished - 3 Nov 2000

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