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The Trimerization Domain of Nemo Is Composed of the Interacting C-terminal CC2 and LZ Coiled-coil Subdomains

Abstract : NEMO (NF-κB essential modulator) plays a key role in the canonical NF-κB pathway as the scaffold/regulatory component of the IκB kinase (IKK) complex. The self-association of NEMO involves the C-terminal halves of the polypeptide chains containing two putative coiled-coil motifs (a CC2 and a LZ leucine zipper), a proline-rich region, and a ZF zinc finger motif. Using purified truncation mutants, we showed that the minimal oligomerization domain of NEMO is the CC2-LZ segment and that both CC2 and LZ subdomains are necessary to restore the LPS-dependent activation of the NF-κB pathway in a NEMO-deficient cell line. We confirmed the association of the oligomerization domain in a trimer and investigated the specific role of CC2 and LZ subdomains in the building of the oligomer. Whereas a recombinant CC2-LZ polypeptide self-associated into a trimer with an association constant close to that of the wild-type protein, the isolated CC2 and LZ peptides, respectively, formed trimers and dimers with weaker association constants. Upon mixing, isolated CC2 and LZ peptides associated to form a stable hetero-hexamer as shown by gel filtration and fluorescence anisotropy experiments. We propose a structural model for the organization of the oligomerization domain of activated NEMO in which three C-terminal domains associate into a pseudo-hexamer forming a six-helix bundle. This model is discussed in relation to the mechanism of activation of the IKK complex by upstream activators.
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Submitted on : Friday, July 2, 2021 - 3:34:40 PM
Last modification on : Tuesday, October 19, 2021 - 10:27:18 PM
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Fabrice Agou, François Traincard, Emilie Vinolo, Gilles Courtois, Shoji Yamaoka, et al.. The Trimerization Domain of Nemo Is Composed of the Interacting C-terminal CC2 and LZ Coiled-coil Subdomains. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2004, 279 (27), pp.27861-27869. ⟨10.1074/jbc.M314278200⟩. ⟨pasteur-03276984⟩

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