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Downton P.♦, Bagnall James S.♦, England H.♦, Spiller David G.♦, Humphreys Neil E.♦, Jackson Dean A.♦, Paszek P., White Michael R.R.♦, Adamson Antony D.♦, Overexpression of IκB⍺ modulates NF-κB activation of inflammatory target gene expression,
Frontiers in Molecular Biosciences, ISSN: 2296-889X, DOI: 10.3389/fmolb.2023.1187187, Vol.10, pp.1187187-1-15, 2023Abstract: Cells respond to inflammatory stimuli such as cytokines by activation of the nuclear factor-κB (NF-κB) signalling pathway, resulting in oscillatory translocation of the transcription factor p65 between nucleus and cytoplasm in some cell types. We investigate the relationship between p65 and inhibitor-κB⍺ (IκBα) protein levels and dynamic properties of the system, and how this interaction impacts on the expression of key inflammatory genes. Using bacterial artificial chromosomes, we developed new cell models of IκB⍺-eGFP protein overexpression in a pseudo-native genomic context. We find that cells with high levels of the negative regulator IκBα remain responsive to inflammatory stimuli and maintain dynamics for both p65 and IκBα. In contrast, canonical target gene expression is dramatically reduced by overexpression of IκBα, but can be partially rescued by overexpression of p65. Treatment with leptomycin B to promote nuclear accumulation of IκB⍺ also suppresses canonical target gene expression, suggesting a mechanism in which nuclear IκB⍺ accumulation prevents productive p65 interaction with promoter binding sites. This causes reduced target promoter binding and gene transcription, which we validate by chromatin immunoprecipitation and in primary cells. Overall, we show how inflammatory gene transcription is modulated by the expression levels of both IκB⍺ and p65. This results in an anti-inflammatory effect on transcription, demonstrating a broad mechanism to modulate the strength of inflammatory response. Keywords: NF-κB, inflammation, IκB⍺, overexpression, gene expression, localisation Affiliations:
Downton P. | - | other affiliation | Bagnall James S. | - | other affiliation | England H. | - | other affiliation | Spiller David G. | - | other affiliation | Humphreys Neil E. | - | other affiliation | Jackson Dean A. | - | other affiliation | Paszek P. | - | IPPT PAN | White Michael R.R. | - | University of Manchester
(GB) | Adamson Antony D. | - | other affiliation |
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