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Since airway epithelial cells respond to airborne insults and potentiate inflammation, IKK\u03b5 expression was characterized in pulmonary epithelial cell lines (A549, BEAS-2B) and primary human bronchial epithelial cells grown as submersion or differentiated air-liquid interface cultures. IKK\u03b5 expression was up-regulated by the pro-inflammatory cytokines, interleukin-1\u03b2 (IL-1\u03b2) and tumour necrosis factor-\u03b1 (TNF\u03b1). Thus, mechanistic interrogations in A549 cells were used to demonstrate the NF-\u03baB dependence of cytokine-induced IKK\u03b5. Furthermore, chromatin immunoprecipitation in A549 and BEAS-2B cells revealed robust recruitment of the NF-\u03baB subunit, p65, to one 5\u2032 and two intronic regions within the IKK\u03b5 locus (IKBKE). In addition, IL-1\u03b2 and TNF\u03b1 induced strong RNA polymerase 2 recruitment to the 5\u2032 region, the first intron, and the transcription start site. Stable transfection of the p65-binding regions into A549 cells revealed IL-1\u03b2- and TNF\u03b1-inducible reporter activity that required NF-\u03baB, but was not repressed by glucocorticoid. While critical NF-\u03baB motifs were identified in the 5\u2032 and downstream intronic regions, the first intronic region did not contain functional NF-\u03baB motifs. Thus, IL-1\u03b2- and TNF\u03b1-induced IKK\u03b5 expression involves three NF-\u03baB-binding regions, containing multiple functional NF-\u03baB motifs, and potentially other mechanisms of p65 binding through non-classical NF-\u03baB binding motifs. By enhancing IKK\u03b5 expression, IL-1\u03b2 may prime, or potentiate, responses to alternative stimuli, as modelled by IKK\u03b5 phosphorylation induced by phorbol 12-myristate 13-acetate. However, since IKK\u03b5 expression was only partially repressed by glucocorticoid, IKK\u03b5-dependent responses could contribute to glucocorticoid-resistant disease.<\/jats:p>","DOI":"10.1042\/bcj20230461","type":"journal-article","created":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T15:31:21Z","timestamp":1719588681000},"page":"959-980","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Inducible gene expression of I\u03baB-kinase \u03b5 is dependent on nuclear factor-\u03baB in human pulmonary epithelial cells"],"prefix":"10.1042","volume":"481","author":[{"given":"Amandah","family":"Necker-Brown","sequence":"first","affiliation":[{"name":"1Department of Physiology and Pharmacology, University of Calgary, Calgary, AB, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cora","family":"Kooi","sequence":"additional","affiliation":[{"name":"1Department of Physiology and Pharmacology, University of Calgary, Calgary, AB, Canada"},{"name":"2Department of Medicine, Lung Health Research Group. 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