PUBLICATION

A Zebrafish Forward Genetic Screen Identifies an Indispensible Threonine Residue in the Kinase Domain of PRKD2

Authors
Giardoglou, P., Bournele, D., Park, M., Kanoni, S., Dedoussis, G.V., Steinberg, S.F., Deloukas, P., Beis, D.
ID
ZDB-PUB-210220-1
Date
2021
Source
Biology Open   10(3): (Journal)
Registered Authors
Beis, Dimitris
Keywords
Cardiac valves, Cardiovascular development, Protein kinase D2, Zebrafish
MeSH Terms
  • Protein Interaction Domains and Motifs*
  • Protein Kinase D2/chemistry
  • Protein Kinase D2/genetics*
  • Protein Kinase D2/metabolism
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Phenotype
  • Amino Acid Sequence
  • Animals
  • Organogenesis/genetics
  • Ectopic Gene Expression
  • Humans
  • Heart/embryology
  • Mutation*
  • Enzyme Activation
  • Threonine/chemistry
  • Threonine/genetics*
  • Amino Acid Substitution
(all 18)
PubMed
33597201 Full text @ Biol. Open
Abstract
Protein Kinase D2 belongs to a family of evolutionarily conserved enzymes regulating several biological processes. In a forward genetic screen for zebrafish cardiovascular mutants, we identified a mutation in the prkd2 gene. Homozygous mutant embryos develop as wild-type up to 36hours post-fertilization and initiate blood flow, but fail to maintain it, resulting in a complete outflow tract stenosis. We identified a mutation in the prkd2 gene that results in a T757A substitution at a conserved residue in the kinase domain activation loop (T714A in human PRKD2) that disrupts catalytic activity and drives this phenotype. Homozygous mutants survive without circulation for several days, allowing us to study the extreme phenotype of no intracardiac flow, in the background of a functional heart. We show dysregulation of atrioventricular and outflow tract markers in the mutants and higher sensitivity to the Calcineurin inhibitor, Cyclosporin A. Finally we identify TBX5 as a potential regulator of PRKD2. Our results implicate PRKD2 catalytic activity in outflow tract development in zebrafish.
Genes / Markers
Figures
Figure Gallery (7 images)
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
s411
    		Point Mutation
    s843TgTransgenic Insertion
      s879TgTransgenic Insertion
        s939TgTransgenic Insertion
          1 - 4 of 4
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          Human Disease / Model
          No data available
          Sequence Targeting Reagents
          Target Reagent Reagent Type
          prkd2MO2-prkd2MRPHLNO
          tbx5aMO5-tbx5aMRPHLNO
          1 - 2 of 2
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          Fish
          No data available
          Antibodies
          Orthology
          No data available
          Engineered Foreign Genes
          Marker Marker Type Name
          DsRedEFGDsRed
          EGFPEFGEGFP
          mCherryEFGmCherry
          1 - 3 of 3
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          Mapping
          No data available
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          Citation
          Giardoglou, P., Bournele, D., Park, M., Kanoni, S., Dedoussis, G.V., Steinberg, S.F., Deloukas, P., Beis, D. (2021) A Zebrafish Forward Genetic Screen Identifies an Indispensible Threonine Residue in the Kinase Domain of PRKD2. Biology Open. 10(3):.