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Sensing of Lys 63-linked polyubiquitination by NEMO is a key event in NF-κB activation

Nature Cell Biology volume 8, pages 398–406 (2006)Cite this article

A Corrigendum to this article was published on 16 April 2006

Abstract

The transcription factor NF-κB is sequestered in the cytoplasm in a complex with IκB1. Almost all NF-κB activation pathways converge on IκB kinase (IKK), which phosphorylates IκB resulting in Lys 48-linked polyubiquitination of IκB and its degradation. This allows migration of NF-κB to the nucleus where it regulates gene expression2. IKK has two catalytic subunits, IKKα and IKKβ, and a regulatory subunit, IKKγ or NEMO. NEMO is essential for NF-κB activation, and NEMO dysfunction in humans is the cause of incontinentia pigmenti and hypohidrotic ectodermal dysplasia and immunodeficiency (HED–ID)3. The recruitment of IKK to occupied cytokine receptors, and its subsequent activation, are dependent on the attachment of Lys 63-linked polyubiquitin chains to signalling intermediates such as receptor-interacting protein (RIP). Here, we show that NEMO binds to Lys 63- but not Lys 48-linked polyubiquitin, and that single point mutations in NEMO that prevent binding to Lys 63-linked polyubiquitin also abrogates the binding of NEMO to RIP in tumour necrosis factor (TNF)-α-stimulated cells, the recruitment of IKK to TNF receptor (TNF-R) 1, and the activation of IKK and NF-κB. RIP is also destabilized in the absence of NEMO binding and undergoes proteasomal degradation in TNF-α-treated cells. These results provide a mechanism for NEMO's critical role in IKK activation, and a key to understanding the link between cytokine-receptor proximal signalling and IKK and NF-κB activation.

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Figure 1: NEMO binds to ubiquitin through a region encompassing the CC2 and leucine zipper domains.
Figure 2: NEMO specifically binds Lys 63-linked polyubiquitin chains.
Figure 3: NEMO mutants that cannot bind polyubiquitin are defective in mediating IKK and NF-κB activation.
Figure 4: NEMO specifically binds Lys 63-polyubiquitinated RIP.
Figure 5: NEMO binding of polyubiquitin recruits IKK to TNF-R1 and protects RIP from TNF-α-induced degradation.

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Acknowledgements

We are grateful to Z. Liu for critical review of this manuscript. S.M.S. is a Kimmel Scholar. This research was supported by the Intramural Research Program of the National Institutes of Health, Center for Cancer Research, National Cancer Institute.

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  1. Laboratory of Immune Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Chuan-Jin Wu, Dietrich B. Conze, Tao Li, Srinivasa M. Srinivasula & Jonathan D. Ashwell

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Correspondence to Jonathan D. Ashwell.

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Wu, CJ., Conze, D., Li, T. et al. Sensing of Lys 63-linked polyubiquitination by NEMO is a key event in NF-κB activation. Nat Cell Biol 8, 398–406 (2006). https://doi.org/10.1038/ncb1384

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