Characterization and expression analysis of Th-POK from the Japanese pufferfish, Takifugu rubripes

Ryusuke Nagamine, Hiroki Korenaga, Masahiro Sakai, Christopher J. Secombes, Tomoya Kono*

*Corresponding author for this work

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

In fish, T cell lineage commitment has not been studied, although there are reports related to CD4 and CD8 positive cells. This study describes the cloning and analysis of a master regulator involved in this process, the Th-POK gene in Japanese pufferfish, Takifugu rubripes. The Fugu Th-POK cDNA was composed of 1901. bp, with a 75. bp 5'-UTR, a 131. bp 3'-UTR, and a 1692. bp open reading frame which translates into a peptide of 564 amino acid residues. The deduced Fugu Th-POK protein contained a BTB/POZ domain, Krüppel motif (H/C linker) and Krüppel-like zinc finger DNA binding domain with C2H2 structure. The homology analysis of Fugu Th-POK (ZBTB7B) with other known ZBTB7 members (ZBTB7A, 7C) showed low identity, and the phylogenetic tree analysis showed the Fugu Th-POK clustered with the mammalian Th-POK, away from other ZBTB7 members. The analysis of transcriptional control region of Th-POK gene suggested that the 5'-flanking region and intron 1 include numerous canonical binding motifs for transcription factors regulating T cell development. The genomic organization of the Fugu Th-POK gene was composed of three exons and two introns, and its structure was identical to that of its human counterpart. Comparison of the Fugu and human genomes showed that high levels of conserved synteny existed around the Th-POK gene. The high expression of the Fugu Th-POK gene in unstimulated tissues was seen in head kidney, muscle, skin and gills. Moreover, the expression of the Fugu Th-POK gene in thymic cells was increased by LPS, polyI:C and PHA stimulation.

Original languageEnglish
Pages (from-to)124-132
Number of pages9
JournalComparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
Volume164
Issue number2
Early online date26 Nov 2012
DOIs
Publication statusPublished - Feb 2013

Fingerprint

Takifugu
Tetraodontiformes
Genes
T-cells
Introns
5' Flanking Region
Cloning
5' Untranslated Regions
Head Kidney
3' Untranslated Regions
Synteny
T-Lymphocytes
Fish
Muscle
Zinc Fingers
Zinc
Exons
Cell Lineage
Skin
Human Genome

Keywords

  • Structural analysis
  • Synteny
  • Takifugu rubripes
  • Th-POK
  • Transcriptional analysis

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

Cite this

Characterization and expression analysis of Th-POK from the Japanese pufferfish, Takifugu rubripes. / Nagamine, Ryusuke; Korenaga, Hiroki; Sakai, Masahiro; Secombes, Christopher J.; Kono, Tomoya.

In: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology, Vol. 164, No. 2, 02.2013, p. 124-132.

Research output: Contribution to journalArticle

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abstract = "In fish, T cell lineage commitment has not been studied, although there are reports related to CD4 and CD8 positive cells. This study describes the cloning and analysis of a master regulator involved in this process, the Th-POK gene in Japanese pufferfish, Takifugu rubripes. The Fugu Th-POK cDNA was composed of 1901. bp, with a 75. bp 5'-UTR, a 131. bp 3'-UTR, and a 1692. bp open reading frame which translates into a peptide of 564 amino acid residues. The deduced Fugu Th-POK protein contained a BTB/POZ domain, Kr{\"u}ppel motif (H/C linker) and Kr{\"u}ppel-like zinc finger DNA binding domain with C2H2 structure. The homology analysis of Fugu Th-POK (ZBTB7B) with other known ZBTB7 members (ZBTB7A, 7C) showed low identity, and the phylogenetic tree analysis showed the Fugu Th-POK clustered with the mammalian Th-POK, away from other ZBTB7 members. The analysis of transcriptional control region of Th-POK gene suggested that the 5'-flanking region and intron 1 include numerous canonical binding motifs for transcription factors regulating T cell development. The genomic organization of the Fugu Th-POK gene was composed of three exons and two introns, and its structure was identical to that of its human counterpart. Comparison of the Fugu and human genomes showed that high levels of conserved synteny existed around the Th-POK gene. The high expression of the Fugu Th-POK gene in unstimulated tissues was seen in head kidney, muscle, skin and gills. Moreover, the expression of the Fugu Th-POK gene in thymic cells was increased by LPS, polyI:C and PHA stimulation.",
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note = "Acknowledgments This work was supported financially by the Program “Improvement of Research Environment for Young Researchers” from the Japanese Ministry of Education, Culture, Sports, Science and Technology, a Grant-in-Aid for Young Scientists (23780199) and a grant for Scientific Research on Priority Areas from the University of Miyazaki.",
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