Abstract
The transcription factor NF-κB plays a pivotal role in innate immunity in response to a variety of stimuli, and the coordinated regulation of this pathway determines the proper host responses to extracellular signals. In this study, we identified RACK1 as a novel negative regulator of NF-κB signaling, NF-κB-mediated cytokine induction and inflammatory reactions. RACK1 physically associates with the IKK complex in a TNF-triggered manner. This interaction interferes with the recruitment of the IKK complex to TRAF2, which is a critical step for IKK phosphorylation and subsequent activation triggered by TNF. By modulating the interaction between TRAF2 and IKK, RACK1 regulates the levels of NF-κB activation in response to different intensities of stimuli. Our findings suggest that RACK1 plays an important role in controlling the sensitivity of TNF-triggered NF-κB signaling by regulating IKK activation and provide new insight into the negative regulation of inflammatory reactions.
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Acknowledgements
We are grateful to Dr Xue-Min Zhang (Academy of military medical sciences, Beijing) for IKK constructs, and to Dr Xiao-Fan Wang (Duke university, Durham) and Dr Dang-Sheng Li for their constructive suggestions on our work. This work was supported by the National Basic Research Program of China (2010CB912102, Ministry of Science and Technology Key Program (2012ZX10002009-017), the National Natural Science Foundation of China (81230058, 30930023, 31100551, 31201046, 81021002), CAS/SAFEA International Partnership Program for Creative Research Teams, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (SIBS2012004), and Technology Commission of Shanghai Municipality (12XD1405600).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
RACK1 repressed the activity of the NF-κB luciferase reporter and the expression of NF-κB target genes. (PDF 105 kb)
Supplementary information, Figure S2
RACK1 inhibited the TNF signaling pathway by influencing the IκBα degradation induced by IKKβ. (PDF 222 kb)
Supplementary information, Figure S3
Binding assays between RACK1 and the IKK complex. (PDF 155 kb)
Supplementary information, Figure S4
Truncated mutants of IKKα and IKKβ that were used in mapping experiments. (PDF 122 kb)
Supplementary information, Figure S5
RACK1 modulated IKK activity through the TNFR1 complex. (PDF 175 kb)
Supplementary information, Figure S6
The expression of RACK1 in 293T and macrophages. (PDF 141 kb)
Supplementary information, Figure S7
Various RACK1 mutants inhibited the TNF-induced NF-κB signaling pathway. (PDF 155 kb)
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Yao, F., Long, LY., Deng, YZ. et al. RACK1 modulates NF-κB activation by interfering with the interaction between TRAF2 and the IKK complex. Cell Res 24, 359–371 (2014). https://doi.org/10.1038/cr.2013.162
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DOI: https://doi.org/10.1038/cr.2013.162
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