PUBLICATION

Nkd1 functions downstream of Axin2 to attenuate Wnt signaling

Authors
Bell, I., Khan, H., Stutt, N., Horn, M., Hydzik, T., Lum, W., Rea, V., Clapham, E., Hoeg, L., Van Raay, T.J.
ID
ZDB-PUB-240425-6
Date
2024
Source
Molecular biology of the cell   35(7): ar93 (Journal)
Registered Authors
Stutt, Nathan, Van Raay, Terry
Keywords
none
MeSH Terms
  • Carrier Proteins
  • Adaptor Proteins, Signal Transducing/genetics
  • Adaptor Proteins, Signal Transducing/metabolism
  • Axin Protein*/genetics
  • Axin Protein*/metabolism
  • Zebrafish*
  • Wnt Signaling Pathway*
  • Zebrafish Proteins*/genetics
  • Zebrafish Proteins*/metabolism
  • beta Catenin*/metabolism
  • Animals
  • Mutation/genetics
(all 12)
PubMed
38656801 Full text @ Mol. Biol. Cell
Abstract
Wnt signaling is a crucial developmental pathway involved in early development as well as stem cell maintenance in adults and its misregulation leads to numerous diseases. Thus, understanding the regulation of this pathway becomes vitally important. Axin2 and Nkd1 are widely utilized negative feedback regulators in Wnt signaling where Axin2 functions to destabilize cytoplasmic β-catenin, and Nkd1 functions to inhibit the nuclear localization of β-catenin. Here, we set out to further understand how Axin2 and Nkd1 regulate Wnt signaling by creating axin2gh1/gh1, nkd1gh2/gh2 single mutants and axin2gh1/gh1;nkd1gh2/gh2 double mutant zebrafish using sgRNA/Cas9. All three Wnt regulator mutants were viable and had impaired heart looping, neuromast migration defects, and behavior abnormalities in common, but there were no signs of synergy in the axin2gh1/gh1;nkd1gh2/gh2 double mutants. Further, Wnt target gene expression by qRT-PCR, and RNA-seq analysis and protein expression by mass spectrometry demonstrated that the double axin2gh1/gh1;nkd1gh2/gh2 mutant resembled the nkd1gh2/gh2 phenotype demonstrating that Nkd1 functions downstream of Axin2. In support of this, the data further demonstrates that Axin2 uniquely alters the properties of β-catenin-dependent transcription having novel readouts of Wnt activity compared to nkd1gh2/gh2 or the axin2gh1/gh1;nkd1gh2/gh2 double mutant. We also tested the sensitivity of the Wnt regulator mutants to exacerbated Wnt signaling, where the single mutants displayed characteristic heightened Wnt sensitivity, resulting in an eyeless phenotype. Surprisingly, this phenotype was rescued in the double mutant, where we speculate that cross-talk between Wnt/β-catenin and Wnt/Planar Cell Polarity pathways could lead to altered Wnt signaling in some scenarios. Collectively, the data emphasizes both the commonality and the complexity in the feedback regulation of Wnt signaling.
Genes / Markers
Figures
Figure Gallery (8 images)
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Expression
No data available
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
gue1
    		Indel
    gue2
      		Small Deletion
      1 - 2 of 2
      Show
      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      axin2CRISPR1-axin2CRISPR
      nkd1CRISPR1-nkd1CRISPR
      1 - 2 of 2
      Show
      Fish
      No data available
      Antibodies
      Name Type Antigen Genes Isotypes Host Organism
      Ab1-actbmonoclonal
        		IgG1Mouse
        Ab2-ctnnbmonoclonalIgG1Mouse
        Ab4-pan-cadherinpolyclonal
          		IgGRabbit
          1 - 3 of 3
          Show
          Orthology
          No data available
          Engineered Foreign Genes
          No data available
          Mapping
          No data available
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          Citation
          Bell, I., Khan, H., Stutt, N., Horn, M., Hydzik, T., Lum, W., Rea, V., Clapham, E., Hoeg, L., Van Raay, T.J. (2024) Nkd1 functions downstream of Axin2 to attenuate Wnt signaling. Molecular biology of the cell. 35(7):ar93.