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SiRNA knockdown screening of possible NF-kB p65 interactors influencing IL-1-NF-kB gene expression

ABSTRACT: The transcription factor NF-κB p65 plays a key role in innate and adaptive immunity. It functions as a master regulator of the inflammatory response by inducing the expression of pro-inflammatory genes including secreted cytokines (IL6), chemokines (IL8, CXCL2), and multiple regulatory intracellular proteins (NFKBIA). NF-κB is activated by the so-called canonical, noncanonical and atypical signalling pathways. In the canonical pathway, activation of the Interleukin-1 receptor complex triggers degradation of the cytosolic p65 inhibitor IBα followed by nuclear translocation of p65 where it is directed by unknown pathways to the enhancers and promoters of pro-inflammatory genes. Despite its pleiotropic functions in adaptive and innate immunity or stress reactions, the entire repertoire of p65 interacting factors that contribute to its general or stimulus-specific functions within the nucleus or other cell compartments is still elusive. We identified the p65 interactomes in intact cells by BioID pulldowns under inflammatory condition (+IL-α). Extensive bioinformatics analysis of the proximity labeling data sets revealed several hundred high confidence interactors including multiple novel factors that have not been reported in the literature. We investigated which of our 38 high-confidence interactors have an impact on IL-1 NF-kB gene expression by siRNA screening. We found 5 interesting hits in the context of inflammation which will be further assessed by microarray analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE244637 | GEO | 2025/03/05

REPOSITORIES: GEO

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