Expression and characterization of a brain-specific protein kinase BSK146 from zebrafish

Chih Ming Chou, Yi Chung Chen, Ming Ting Lee, Gen Der Chen, I. Ching Lu, Shui Tsung Chen, Chang Jen Huang

研究成果: 雜誌貢獻文章

7 引文 (Scopus)

摘要

We have previously identified a novel protein kinase, pk146, in the brain of Tetraodon. In the present study, we cloned the homologous protein kinase gene encoding a protein of 385 amino acid residues from zebrafish. The overall amino acid sequence and the kinase domain of zebrafish BSK146 shows 48% and 69% identity to that of rat sbk, a SH3-containing serine/threonine protein kinase. By whole-mount in situ hybridization and RT-PCR, the expression of bsk146 mRNA was mainly in the brain. To explore the in vivo function of BSK146 during zebrafish development, we used morpholino knockdown approach and found that BSK146 morphants displayed enlarged hindbrain ventricle and smaller eyes. Whole-mount in situ hybridization was further performed to analyze the brain defects in BSK146-MO-injected embryos. The expression of brain-specific markers, such as otx2, pax2.1, and krox20, was found normal in morphant embryos at 24 hpf, while expression of pax2.1 exerted changes in midbrain-hindbrain boundary and hindbrain in morphant embryos at 48 hpf. These data suggest that BSK146 may play an important role in later ventricle expansion in zebrafish brain development. Although the recombinant BSK146 protein produced in insect cells was active and could phosphorylate both histone H1 and histone 2B, the endogenous substrate of BSK146 in the embryonic brain of zebrafish is not clear at the present time and needs further investigation.
原文英語
頁(從 - 到)767-775
頁數9
期刊Biochemical and Biophysical Research Communications
340
發行號3
DOIs
出版狀態已發佈 - 二月 17 2006

指紋

EphA5 Receptor
Zebrafish
Protein Kinases
Brain
Rhombencephalon
Embryonic Structures
Histones
In Situ Hybridization
Amino Acids
Recombinant proteins
Morpholinos
Gene encoding
Protein-Serine-Threonine Kinases
Mesencephalon
Recombinant Proteins
Insects
Rats
Amino Acid Sequence
Phosphotransferases
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

引用此文

Expression and characterization of a brain-specific protein kinase BSK146 from zebrafish. / Chou, Chih Ming; Chen, Yi Chung; Lee, Ming Ting; Chen, Gen Der; Lu, I. Ching; Chen, Shui Tsung; Huang, Chang Jen.

於: Biochemical and Biophysical Research Communications, 卷 340, 編號 3, 17.02.2006, p. 767-775.

研究成果: 雜誌貢獻文章

Chou, Chih Ming ; Chen, Yi Chung ; Lee, Ming Ting ; Chen, Gen Der ; Lu, I. Ching ; Chen, Shui Tsung ; Huang, Chang Jen. / Expression and characterization of a brain-specific protein kinase BSK146 from zebrafish. 於: Biochemical and Biophysical Research Communications. 2006 ; 卷 340, 編號 3. 頁 767-775.
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abstract = "We have previously identified a novel protein kinase, pk146, in the brain of Tetraodon. In the present study, we cloned the homologous protein kinase gene encoding a protein of 385 amino acid residues from zebrafish. The overall amino acid sequence and the kinase domain of zebrafish BSK146 shows 48{\%} and 69{\%} identity to that of rat sbk, a SH3-containing serine/threonine protein kinase. By whole-mount in situ hybridization and RT-PCR, the expression of bsk146 mRNA was mainly in the brain. To explore the in vivo function of BSK146 during zebrafish development, we used morpholino knockdown approach and found that BSK146 morphants displayed enlarged hindbrain ventricle and smaller eyes. Whole-mount in situ hybridization was further performed to analyze the brain defects in BSK146-MO-injected embryos. The expression of brain-specific markers, such as otx2, pax2.1, and krox20, was found normal in morphant embryos at 24 hpf, while expression of pax2.1 exerted changes in midbrain-hindbrain boundary and hindbrain in morphant embryos at 48 hpf. These data suggest that BSK146 may play an important role in later ventricle expansion in zebrafish brain development. Although the recombinant BSK146 protein produced in insect cells was active and could phosphorylate both histone H1 and histone 2B, the endogenous substrate of BSK146 in the embryonic brain of zebrafish is not clear at the present time and needs further investigation.",
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AU - Huang, Chang Jen

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AB - We have previously identified a novel protein kinase, pk146, in the brain of Tetraodon. In the present study, we cloned the homologous protein kinase gene encoding a protein of 385 amino acid residues from zebrafish. The overall amino acid sequence and the kinase domain of zebrafish BSK146 shows 48% and 69% identity to that of rat sbk, a SH3-containing serine/threonine protein kinase. By whole-mount in situ hybridization and RT-PCR, the expression of bsk146 mRNA was mainly in the brain. To explore the in vivo function of BSK146 during zebrafish development, we used morpholino knockdown approach and found that BSK146 morphants displayed enlarged hindbrain ventricle and smaller eyes. Whole-mount in situ hybridization was further performed to analyze the brain defects in BSK146-MO-injected embryos. The expression of brain-specific markers, such as otx2, pax2.1, and krox20, was found normal in morphant embryos at 24 hpf, while expression of pax2.1 exerted changes in midbrain-hindbrain boundary and hindbrain in morphant embryos at 48 hpf. These data suggest that BSK146 may play an important role in later ventricle expansion in zebrafish brain development. Although the recombinant BSK146 protein produced in insect cells was active and could phosphorylate both histone H1 and histone 2B, the endogenous substrate of BSK146 in the embryonic brain of zebrafish is not clear at the present time and needs further investigation.

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