Linked Life Data

19527677

Source:http://linkedlifedata.com/resource/pubmed/id/19527677

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2009-8-3
pubmed:databankReference
pubmed:abstractText
Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKP) dephosphorylates and regulates multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaMKs). However, the biological functions of this enzyme have not been clarified in vivo. To investigate the biological significance of CaMKP during zebrafish embryogenesis, we cloned and characterized zebrafish CaMKP (zCaMKP). The isolated cDNA clone possessed an open reading frame of 1272bp encoding 424 amino acids and shared 47% and 48% amino acid identity with rat and human CaMKP, respectively. Interestingly, zCaMKP lacks the Glu cluster corresponding to residues 101-109 in the rat enzyme, and was not activated by polycations such as poly-l-lysine. The recombinant zCaMKP required Mg(2+) rather than Mn(2+) for activity. Furthermore, zCaMKP dephosphorylated CaMKIV but not phosphorylase a, alpha-casein, or extracellular signal-regulating kinase (ERK), suggesting that the enzyme regulates Ca(2+) signaling pathways in zebrafish. Cotransfection of zCaMKP with mammalian CaMKI significantly decreased phospho-CaMKI in ionomycin-stimulated 293T cells. During embryogenesis, the expression of zCaMKP increased gradually after 48h post-fertilization, as demonstrated by Western blotting using an anti-zCaMKP antibody. The knockdown of the zCaMKP gene with morpholino-based antisense oligonucleotides resulted in an increased incidence of embryos with severe morphological and cellular abnormalities, i.e., a significant increase in the number of round-shaped embryos and apoptotic cells in the whole body. A marked decrease in zCaMKP expression was observed in the antisense- but not control oligo-injected embryos. Embryonic death was rescued by coinjection with recombinant rat CaMKP but not with phosphatase-dead mutant (D194A). These results clearly show the significance of zCaMKP during zebrafish embryogenesis.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1096-0384
pubmed:author
pubmed:issnType
Electronic
pubmed:day
1
pubmed:volume
488
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
48-59
pubmed:meshHeading
pubmed-meshheading:19527677-Amino Acid Sequence, pubmed-meshheading:19527677-Animals, pubmed-meshheading:19527677-Base Sequence, pubmed-meshheading:19527677-Calcium-Calmodulin-Dependent Protein Kinases, pubmed-meshheading:19527677-Cell Line, pubmed-meshheading:19527677-Cell Survival, pubmed-meshheading:19527677-Cloning, Molecular, pubmed-meshheading:19527677-DNA, Complementary, pubmed-meshheading:19527677-Embryonic Development, pubmed-meshheading:19527677-Escherichia coli, pubmed-meshheading:19527677-Gene Expression Regulation, Developmental, pubmed-meshheading:19527677-Gene Knockdown Techniques, pubmed-meshheading:19527677-Humans, pubmed-meshheading:19527677-Intracellular Space, pubmed-meshheading:19527677-Molecular Sequence Data, pubmed-meshheading:19527677-Phosphoprotein Phosphatases, pubmed-meshheading:19527677-Phosphorylation, pubmed-meshheading:19527677-Protein Transport, pubmed-meshheading:19527677-Rats, pubmed-meshheading:19527677-Recombinant Proteins, pubmed-meshheading:19527677-Substrate Specificity, pubmed-meshheading:19527677-Zebrafish, pubmed-meshheading:19527677-Zebrafish Proteins
pubmed:year
2009
pubmed:articleTitle
Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKP) is indispensable for normal embryogenesis in zebrafish, Danio rerio.
pubmed:affiliation
Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, Japan. sueyoshi@ag.kagawa-u.ac.jp
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't