Expression and gene knockdown of zebrafish Ca2+/calmodulin-dependent protein kinase Idelta-LL
- Authors
- Senga, Y., Yoshioka, K., Kameshita, I., and Sueyoshi, N.
- ID
- ZDB-PUB-131112-5
- Date
- 2013
- Source
- Archives of biochemistry and biophysics 540(1-2): 41-52 (Journal)
- Registered Authors
- Keywords
- CaMKlδ delta, embryogenesis, gene knockdown, protein phosphorylation, zebrafish
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Calcium-Calmodulin-Dependent Protein Kinase Type 1/chemistry
- Calcium-Calmodulin-Dependent Protein Kinase Type 1/deficiency*
- Calcium-Calmodulin-Dependent Protein Kinase Type 1/genetics*
- Calcium-Calmodulin-Dependent Protein Kinase Type 1/metabolism
- Cloning, Molecular
- DNA, Complementary/genetics
- Gene Expression Regulation, Enzymologic
- Gene Knockdown Techniques*
- Intracellular Space/metabolism
- Molecular Sequence Data
- Protein Isoforms/chemistry
- Protein Isoforms/deficiency
- Protein Isoforms/genetics
- Protein Isoforms/metabolism
- Protein Transport
- Zebrafish/genetics*
- PubMed
- 24099663 Full text @ Arch. Biochem. Biophys.
Ca2+/calmodulin-dependent protein kinase Iδ (CaMKIδ) is expressed ubiquitously, but little is known about its physiological functions. Recently, we cloned and characterized two splice variants of zebrafish (Danio rerio) CaMKIδ (CaMKIδ-S/L). In the present study we cloned a new CaMKIδ isoform, CaMKIδ-LL, encoded by a different gene from CaMKIδ-S/L. While the catalytic domain of CaMKIδ-LL showed 86% identity that of CaMKIδ-S/L, it had a unique C-terminal sequence. To clarify the functional role of CaMKIδ-LL, we investigated the biological significance of this new isoform during zebrafish embryogenesis. Although CaMKIδ-LL exhibited essentially the same catalytic properties and substrate specificities as the other CaMKIδ isoforms, it showed different temporal and spatial expression. During zebrafish embryogenesis, RT-PCR analysis detected CaMKIδ-LL expression after 48 h post-fertilization. Western blotting in adult zebrafish demonstrated that CaMKIδ-LL is expressed in the brain, the eye, and, abundantly, in fins. Knockdown of CaMKIδ-LL expression using morpholino-based antisense oligonucleotides resulted in an increase in abnormal embryos with small fins and underdeveloped cartilage. These phenotypes were rescued by co-injection with recombinant CaMKIδ-LL. These results clearly indicated that CaMKIδ-LL plays an important role in the generation of cartilage and fins during zebrafish embryogenesis.