PUBLICATION

NCK-associated protein 1 like (nckap1l) minor splice variant regulates intrahepatic biliary network morphogenesis

Authors
Ghaffari, K., Pierce, L.X., Roufaeil, M., Gibson, I., Tae, K., Sahoo, S., Cantrell, J.R., Andersson, O., Lau, J., Sakaguchi, T.F.
ID
ZDB-PUB-210320-10
Date
2021
Source
PLoS Genetics   17: e1009402 (Journal)
Registered Authors
Pierce, Lain, Sakaguchi, Takuya
Keywords
none
MeSH Terms
  • Adaptor Proteins, Signal Transducing/genetics*
  • Alleles
  • Alternative Splicing*
  • Animals
  • Animals, Genetically Modified
  • Bile Ducts, Intrahepatic/embryology*
  • Bile Ducts, Intrahepatic/metabolism*
  • Cyclin-Dependent Kinase 5/genetics
  • Cyclin-Dependent Kinase 5/metabolism
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Gene Order
  • Genetic Testing
  • Genetic Variation
  • Liver/metabolism
  • Models, Biological
  • Morphogenesis/genetics*
  • Mutation
  • Phenotype
  • RNA Isoforms
  • Zebrafish
  • rac1 GTP-Binding Protein/genetics
  • rac1 GTP-Binding Protein/metabolism
PubMed
33739979 Full text @ PLoS Genet.
Abstract
Impaired formation of the intrahepatic biliary network leads to cholestatic liver diseases, which are frequently associated with autoimmune disorders. Using a chemical mutagenesis strategy in zebrafish combined with computational network analysis, we screened for novel genes involved in intrahepatic biliary network formation. We positionally cloned a mutation in the nckap1l gene, which encodes a cytoplasmic adaptor protein for the WAVE regulatory complex. The mutation is located in the last exon after the stop codon of the primary splice isoform, only disrupting a previously unannotated minor splice isoform, which indicates that the minor splice isoform is responsible for the intrahepatic biliary network phenotype. CRISPR/Cas9-mediated nckap1l deletion, which disrupts both the primary and minor isoforms, showed the same defects. In the liver of nckap1l mutant larvae, WAVE regulatory complex component proteins are degraded specifically in biliary epithelial cells, which line the intrahepatic biliary network, thus disrupting the actin organization of these cells. We further show that nckap1l genetically interacts with the Cdk5 pathway in biliary epithelial cells. These data together indicate that although nckap1l was previously considered to be a hematopoietic cell lineage-specific protein, its minor splice isoform acts in biliary epithelial cells to regulate intrahepatic biliary network formation.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping