Noncanonical NF-κB Activation by the Oncoprotein Tio Occurs Through a Nonconserved TRAF3-Binding Motif
Preventing NF-κB Pathway Crosstalk
The transcription factor NF-κB (nuclear factor κB) can be activated by so-called canonical and noncanonical pathways. Activation of the noncanonical NF-κB pathway blocks the constitutive degradation of the kinase NIK (NF-κB–inducing kinase), which leads to the generation of an NF-κB subunit required for target gene expression. The viral oncoprotein Tio mimics a constitutively active receptor upstream of NF-κB signaling, and de Jong et al. found that it contains a binding motif not conserved in other proteins that bind to TRAF3 (tumor necrosis factor receptor–associated factor 3), an inhibitor of noncanonical NF-κB signaling. This TRAF3-binding motif enabled Tio to specifically activate noncanonical NF-κB signaling without triggering crosstalk with the canonical pathway. Tio signaling did not result in TRAF3 degradation; rather, it induced the sequestration of a TRAF3-containing degradative complex from NIK to stimulate the noncanonical pathway. These data suggest that Tio might be used as a tool to examine the specific activation of genes targeted by noncanonical NF-κB signaling in the context of viral transformation.
Abstract
Members of the nuclear factor κB (NF-κB) family of transcription factors regulate many cellular functions. Activation of NF-κB signaling is commonly classified as occurring through canonical or noncanonical pathways. Most NF-κB–inducing stimuli, including the viral oncoprotein Tio, lead to a concerted activation of both NF-κB pathways; however, extensive crosstalk at multiple levels between these signaling cascades restricts the ability to discriminate between the canonical and the noncanonical effects. We showed that noncanonical NF-κB activation by Tio depends on a distinct sequence motif that directly recruits tumor necrosis factor receptor–associated factor 3 (TRAF3). Through its TRAF3-binding motif, Tio triggered a ubiquitin-independent depletion of TRAF3 from the cytosol, which prevented TRAF3 from inhibiting signaling through the noncanonical NF-κB cascade. Furthermore, the Tio-TRAF3 interaction did not affect components of the canonical NF-κB signaling pathway or the expression of target genes; thus, Tio induced noncanonical NF-κB independently of crosstalk with the canonical pathway. Together, these data identify a distinct molecular mechanism of noncanonical NF-κB activation that should enable studies into the particular functions of this pathway.
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Supplementary Material
Summary
Fig. S1. Quantification of the extent of processing of p100 to p52 in the presence of Tio deletion mutants.
Fig. S2. Tio does not form complexes with NIK.
Fig. S3. Quantification of TRAF3 protein abundance in the presence of Tio.
Fig. S4. Tio does not use ubiquitination, lysosomal degradation, or cIAP1 or cIAP2 activity to reduce the abundance of TRAF3.
Fig. S5. Quantification of the extent of processing of p100 to p52 in the presence of combinatorial Tio mutants.
Table S1. Primer sequences.
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Information & Authors
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Published In
Science Signaling
Volume 6 | Issue 272
April 2013
April 2013
Copyright
Copyright © 2013, American Association for the Advancement of Science.
Submission history
Received: 14 June 2012
Accepted: 5 April 2013
Acknowledgments
We thank M. Schmidt and I. Müller-Fleckenstein for transformed cell cultures and B. Fleckenstein for valuable suggestions. LBW242 was a gift of Novartis Pharma AG (Basel). Funding: This work was supported by grants from the German Research Foundation (BI465/5-1, GRK1071/C3) to B.B., the Elite Network of Bavaria (BIGSS) and the Friedrich-Alexander-Universität Erlangen-Nürnberg (FFL) to S.J.d.J., SFB796 (project A2) to H.S., and SFB 684 (project A20) and the LifeScience Foundation to A.K. Author contributions: S.J.d.J., A.K., H.S., and B.B. planned the experiments; S.J.d.J., J.-C.A., F.G., H.S., and B.B. performed the experiments; and S.J.d.J., H.S., and B.B. wrote the manuscript. Competing interests: The authors declare that they have no competing interests.
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