title: IκB kinase β (IKKβ) does not mediate feedback inhibition of the insulin signalling cascade.
abstract: In the present study, we have examined whether IKKβ [IκB (inhibitor of nuclear factor κB) kinase β] plays a role in feedback inhibition of the insulin signalling cascade. Insulin induces the phosphorylation of IKKβ, in vitro and in vivo, and this effect is dependent on intact signalling via PI3K (phosphoinositide 3-kinase), but not PKB (protein kinase B). To test the hypothesis that insulin activates IKKβ as a means of negative feedback, we employed a variety of experimental approaches. First, pharmacological inhibition of IKKβ via BMS-345541 did not potentiate insulin-induced IRS1 (insulin receptor substrate 1) tyrosine phosphorylation, PKB phosphorylation or 2-deoxyglucose uptake in differentiated 3T3-L1 adipocytes. BMS-345541 did not prevent insulin-induced IRS1 serine phosphorylation on known IKKβ target sites. Secondly, adenovirus-mediated overexpression of wild-type IKKβ in differentiated 3T3-L1 adipocytes did not suppress insulin-stimulated 2-deoxyglucose uptake, IRS1 tyrosine phosphorylation, IRS1 association with the p85 regulatory subunit of PI3K or PKB phosphorylation. Thirdly, insulin signalling was not potentiated in mouse embryonic fibroblasts lacking IKKβ. Finally, insulin treatment of 3T3-L1 adipocytes did not promote the recruitment of IKKβ to IRS1, supporting our findings that IKKβ, although activated by insulin, does not promote direct serine phosphorylation of IRS1 and does not contribute to the feedback inhibition of the insulin signalling cascade.
authors.affiliation: Cellular and Molecular Metabolism Laboratory, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia. graeme.lancaster@bakeridi.edu.au
authors.forename: Graeme I
authors.initials: GI
authors.lastname: Lancaster
authors.forename: Beata
authors.initials: B
authors.lastname: Skiba
authors.forename: Christine
authors.initials: C
authors.lastname: Yang
authors.forename: Hayley T
authors.initials: HT
authors.lastname: Nicholls
authors.forename: Katherine G
authors.initials: KG
authors.lastname: Langley
authors.forename: M H Stanley
authors.initials: MH
authors.lastname: Chan
authors.forename: Clinton R
authors.initials: CR
authors.lastname: Bruce
authors.forename: Gordon W
authors.initials: GW
authors.lastname: Rewcastle
authors.forename: Peter R
authors.initials: PR
authors.lastname: Shepherd
authors.forename: Michael
authors.initials: M
authors.lastname: Karin
authors.forename: Mark A
authors.initials: MA
authors.lastname: Febbraio
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism; D000818:Animals; D025461:Feedback, Physiological / Q000502:physiology*; D051550:I-kappa B Kinase / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D055504:Insulin Receptor Substrate Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D012694:Serine / Q000235:genetics; D015398:Signal Transduction
chemical_list: D007328:Insulin; D055504:Insulin Receptor Substrate Proteins; D012694:Serine; D051550:I-kappa B Kinase