Novel mechanism of Wnt signalling inhibition mediated by Dickkopf-1 interaction with LRP6/Arrow

Abstract

Wnt signalling has an important role in cell fate determination, tissue patterning and tumorigenesis^1,2,3,4. Secreted antagonists of Wnt include Frizzled (Fz)-related proteins (FRPs)^5,6,7, Cerberus⁸, Wnt inhibitory factor (WIF)⁹ and Dickkopf (Dkk)^10,11. FRPs, Cerberus and WIF have all been shown to act by binding and sequestering Wnt. We report a novel mechanism of Wnt-signalling inhibition by human Dkk-1. Dkk-1 demonstrated no interaction with Wnt but bound a single cell surface site with high affinity (K_D = 0.39 nM). Its receptor was detectable in a complex with a relative molecular mass of 240,000 (M_r 240K) with [¹²⁵I] Dkk-1 by covalent affinity cross-linking. Wnt signalling through β-catenin is mediated by the Fz receptor¹² and a recently identified low-density-lipoprotein-receptor-related co-receptor, LRP6/Arrow^13,14,15. Overproduction of the 200K LRP6 protein, but not of Fz, strikingly increased Dkk-1 binding as well as the amount of the 240K cross-linked complex, which was shown to be composed of Dkk-1 and LRP6. Moreover, Dkk-1 function was completely independent of Fz but LRP6 dramatically interfered with the Dkk-1 inhibition of Wnt signalling. Thus, unlike Wnt antagonists, which exert their effects by molecular mimicry of Fz^5,6,7 or Wnt sequestration through other mechanisms^8,9, Dkk-1 specifically inhibits canonical Wnt signalling by binding to the LRP6 component of the receptor complex.