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
-
- Amino Acid Sequence
- Amino Acid Substitution
- Animals
- Ectopic Gene Expression
- Enzyme Activation
- Heart/embryology
- Humans
- Mutation*
- Organogenesis/genetics
- Phenotype
- Protein Interaction Domains and Motifs*
- Protein Kinase D2/chemistry
- Protein Kinase D2/genetics*
- Protein Kinase D2/metabolism
- Threonine/chemistry
- Threonine/genetics*
- Zebrafish/genetics*
- Zebrafish/metabolism
- PubMed
- 33597201 Full text @ Biol. Open
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):.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping