Endocytosis-Independent Function of Clathrin Heavy Chain in the Control of Basal NF-κB Activation 英文参考文献.docVIP

Endocytosis-IndependentFunctionofClathrinHeavy ChainintheControlofBasalNF-k BActivation ManLyangKim,IsabelSorg,Ce′cileArrieumerlou* Biozentrum,UniversityofBasel,Basel,Switzerland Abstract Background:Nuclearfactor-kB(NF-kB)isatranscriptionfactorthatregulatesthetranscriptionofgenesinvolvedinavariety of biological processes, including innate and adaptive immunity, stress responses and cell proliferation. Constitutive or excessive NF-kB activity has been associated with inflammatory disorders and higher risk of cancer. In contrast to the mechanismscontrollinginducibleactivation,theregulationofbasalNF-kBactivationisnotwellunderstood.Herewetest whetherclathrinheavychain(CHC)contributestotheregulationofbasalNF-kBactivityinepithelialcells. Methodology:UsingRNAinterferencetoreduceendogenousCHCexpression,wefoundthatCHCisrequiredtoprevent constitutive activation of NF-kB and gene expression. Immunofluorescence staining showed constitutive nuclear localization of the NF-kB subunit p65 in absence of stimulation after CHC knockdown. Elevated basal p65 nuclear localizationiscausedbyconstitutivephosphorylationanddegradationofinhibitorofNF-kBalpha(IkBa)throughanIkB kinasea(IKKa)-dependentmechanism.TheroleofCHCinNF-kBsignalingisfunctionallyrelevantasconstitutiveexpression of the proinflammatory chemokine interleukin-8 (IL-8), whose expression is regulated by NF-kB, was found after CHC knockdown. Disruption of clathrin-mediated endocytosis by chemical inhibition or depletion of the m2-subunit of the endocytosis adaptor protein AP-2, and knockdown of clathrin light chain a (CHLa), failed to induce constitutive NF-kB activationandIL-8expression,showingthatCHCactsonNF-kBindependentlyofendocytosisandCLCa. Conclusions: We conclude that CHC functions as a built-in molecular brake that ensures a tight control of basal NF-kB activationandgeneexpressioninunstimulatedcells.Furthermore,ourdatasuggestapotentiallinkbetweenadefectin CHCexpressionandchronicinflammationdisorderandcancer. Citation: