PUBLICATION
Knockdown of nuclear Ca(2+)/calmodulin-dependent protein kinase phosphatase causes developmental abnormalities in zebrafish
- Authors
- Nimura, T., Sueyoshi, N., Ishida, A., Yoshimura, Y., Ito, M., Tokumitsu, H., Shigeri, Y., Nozaki, N., and Kameshita, I.
- ID
- ZDB-PUB-061229-2
- Date
- 2007
- Source
- Archives of biochemistry and biophysics 457(2): 205-216 (Journal)
- Registered Authors
- Yoshimura, Yukihiro
- Keywords
- Zebrafish, CaM kinase, Apoptosis, Phosphatase, CaMKP, CaMKP-N
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Apoptosis
- Base Sequence
- Calcium-Calmodulin-Dependent Protein Kinases/metabolism*
- Cell Line, Tumor
- Cell Nucleus/metabolism
- Embryo, Nonmammalian/metabolism
- Ionomycin/pharmacology
- Ionophores/pharmacology
- Mice
- Molecular Sequence Data
- Mutation
- Nuclear Localization Signals
- Phosphoprotein Phosphatases/genetics
- Phosphoprotein Phosphatases/metabolism*
- Phosphorylation
- Rats
- Zebrafish/abnormalities
- Zebrafish/embryology*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 17169323 Full text @ Arch. Biochem. Biophys.
Citation
Nimura, T., Sueyoshi, N., Ishida, A., Yoshimura, Y., Ito, M., Tokumitsu, H., Shigeri, Y., Nozaki, N., and Kameshita, I. (2007) Knockdown of nuclear Ca(2+)/calmodulin-dependent protein kinase phosphatase causes developmental abnormalities in zebrafish. Archives of biochemistry and biophysics. 457(2):205-216.
Abstract
Nuclear Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKP-N) is an enzyme that dephosphorylates and concomitantly downregulates multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaMKs) in vitro. However, the functional roles of this enzyme in vivo are not well understood. To investigate the biological significance of CaMKP-N during zebrafish embryogenesis, we cloned and characterized zebrafish CaMKP-N (zCaMKP-N). Based on the nucleotide sequences in the zebrafish whole genome shotgun database, we isolated a cDNA clone for zCaMKP-N, which encoded a protein of 633 amino acid residues. Transiently expressed full-length zCaMKP-N in mouse neuroblastoma, Neuro2a cells, was found to be localized in the nucleus. In contrast, the C-terminal truncated mutant lacking RKKRRLDVLPLRR (residues 575-587) had cytoplasmic staining, suggesting that the nuclear localization signal of zCaMKP-N exists in the C-terminal region. Ionomycin treatment of CaMKIV-transfected Neuro2a cells resulted in a marked increase in the phosphorylated form of CaMKIV. However, cotransfection with zCaMKP-N significantly decreased phospho-CaMKIV in ionomycin-stimulated cells. Whole mount in situ hybridization analysis of zebrafish embryos showed that zCaMKP-N is exclusively expressed in the head and neural tube regions. Gene knockdown of zCaMKP-N using morpholino-based antisense oligonucleotides induced significant morphological abnormalities in zebrafish embryos. A number of apoptotic cells were observed in brain and spinal cord of the abnormal embryos. These results suggest that zCaMKP-N plays a crucial role in the early development of zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping