Wnt5a promotes Frizzled-4 signalosome assembly by stabilizing cysteine-rich domain dimerization

Zachary J. DeBruine, Jiyuan Ke, Kaleeckal G. Harikumar, Xin Gu, Peter Borowsky, Bart O. Williams, Wenqing Xu, Laurence J Miller, H. Eric Xu, Karsten Melcher

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Wnt/β-catenin signaling is activated when extracellular Wnt ligands bind Frizzled (FZD) receptors at the cell membrane. Wnts bind FZD cysteine-rich domains (CRDs) with high affinity through a palmitoylated N-terminal “thumb” and a disulfide-stabilized C-terminal “index finger,” yet how these binding events trigger receptor activation and intracellular signaling remains unclear. Here we report the crystal structure of the Frizzled-4 (FZD4) CRD in complex with palmitoleic acid, which reveals a CRD tetramer consisting of two cross-braced CRD dimers. Each dimer is stabilized by interactions of one hydrophobic palmitoleic acid tail with two CRD palmitoleoyl-binding grooves oriented end to end, suggesting that the Wnt palmitoleoyl group stimulates CRD–CRD interaction. Using bioluminescence resonance energy transfer (BRET) in live cells, we show that WNT5A stimulates dimerization of membrane-anchored FZD4 CRDs and oligomerization of full-length FZD4, which requires the integrity of CRD palmitoleoyl-binding residues. These results suggest that FZD receptors may form signalosomes in response to Wnt binding through the CRDs and that the Wnt palmitoleoyl group is important in promoting these interactions. These results complement our understanding of lipoprotein receptor-related proteins 5 and 6 (LRP5/6), Dishevelled, and Axin signalosome assembly and provide a more complete model for Wnt signalosome assembly both intracellularly and at the membrane.

Original languageEnglish (US)
Pages (from-to)916-926
Number of pages11
JournalGenes and Development
Volume31
Issue number9
DOIs
StatePublished - 2017

Fingerprint

Dimerization
Cysteine
Frizzled Receptors
Low Density Lipoprotein Receptor-Related Protein-6
Low Density Lipoprotein Receptor-Related Protein-5
Catenins
Membranes
Energy Transfer
Thumb
Hydrophobic and Hydrophilic Interactions
Disulfides
Fingers
Cell Membrane
Ligands

Keywords

  • Cysteine-rich domain
  • Frizzled
  • Frizzled-4
  • Signalosome
  • Wnt
  • WNT5A

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

Cite this

DeBruine, Z. J., Ke, J., Harikumar, K. G., Gu, X., Borowsky, P., Williams, B. O., ... Melcher, K. (2017). Wnt5a promotes Frizzled-4 signalosome assembly by stabilizing cysteine-rich domain dimerization. Genes and Development, 31(9), 916-926. https://doi.org/10.1101/gad.298331.117

Wnt5a promotes Frizzled-4 signalosome assembly by stabilizing cysteine-rich domain dimerization. / DeBruine, Zachary J.; Ke, Jiyuan; Harikumar, Kaleeckal G.; Gu, Xin; Borowsky, Peter; Williams, Bart O.; Xu, Wenqing; Miller, Laurence J; Xu, H. Eric; Melcher, Karsten.

In: Genes and Development, Vol. 31, No. 9, 2017, p. 916-926.

Research output: Contribution to journalArticle

DeBruine, ZJ, Ke, J, Harikumar, KG, Gu, X, Borowsky, P, Williams, BO, Xu, W, Miller, LJ, Xu, HE & Melcher, K 2017, 'Wnt5a promotes Frizzled-4 signalosome assembly by stabilizing cysteine-rich domain dimerization', Genes and Development, vol. 31, no. 9, pp. 916-926. https://doi.org/10.1101/gad.298331.117
DeBruine, Zachary J. ; Ke, Jiyuan ; Harikumar, Kaleeckal G. ; Gu, Xin ; Borowsky, Peter ; Williams, Bart O. ; Xu, Wenqing ; Miller, Laurence J ; Xu, H. Eric ; Melcher, Karsten. / Wnt5a promotes Frizzled-4 signalosome assembly by stabilizing cysteine-rich domain dimerization. In: Genes and Development. 2017 ; Vol. 31, No. 9. pp. 916-926.
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AU - Gu, Xin

AU - Borowsky, Peter

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AU - Xu, Wenqing

AU - Miller, Laurence J

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AB - Wnt/β-catenin signaling is activated when extracellular Wnt ligands bind Frizzled (FZD) receptors at the cell membrane. Wnts bind FZD cysteine-rich domains (CRDs) with high affinity through a palmitoylated N-terminal “thumb” and a disulfide-stabilized C-terminal “index finger,” yet how these binding events trigger receptor activation and intracellular signaling remains unclear. Here we report the crystal structure of the Frizzled-4 (FZD4) CRD in complex with palmitoleic acid, which reveals a CRD tetramer consisting of two cross-braced CRD dimers. Each dimer is stabilized by interactions of one hydrophobic palmitoleic acid tail with two CRD palmitoleoyl-binding grooves oriented end to end, suggesting that the Wnt palmitoleoyl group stimulates CRD–CRD interaction. Using bioluminescence resonance energy transfer (BRET) in live cells, we show that WNT5A stimulates dimerization of membrane-anchored FZD4 CRDs and oligomerization of full-length FZD4, which requires the integrity of CRD palmitoleoyl-binding residues. These results suggest that FZD receptors may form signalosomes in response to Wnt binding through the CRDs and that the Wnt palmitoleoyl group is important in promoting these interactions. These results complement our understanding of lipoprotein receptor-related proteins 5 and 6 (LRP5/6), Dishevelled, and Axin signalosome assembly and provide a more complete model for Wnt signalosome assembly both intracellularly and at the membrane.

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