Caspase 3 from rock bream (Oplegnathus fasciatus): Genomic characterization and transcriptional profiling upon bacterial and viral inductions

Don Anushka Sandaruwan Elvitigala, Ilson Whang, H. K A Premachandra, Navaneethaiyer Umasuthan, Myung Joo Oh, Sung Ju Jung, Sang Yeob Yeo, Bong Soo Lim, Jeong Ho Lee, Hae Chul Park, Jehee Lee

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Caspase 3 is a prominent mediator of apoptosis and participates in the cell death signaling cascade. In this study, caspase 3 was identified (Rbcasp3) and characterized from rock bream (Oplegnathus fasciatus). The full-length cDNA of Rbcasp3 is 2683 bp and contains an open reading frame of 849 bp, which encodes a 283 amino acid protein with a calculated molecular mass of 31.2 kDa and isoelectric point of 6.31. The amino acid sequence resembles the conventional caspase 3 domain architecture, including crucial amino acid residues in the catalytic site and binding pocket. The genomic length of Rbcasp3 is 7529 bp, and encompasses six exons interrupted by five introns. Phylogenetic analysis affirmed that Rbcasp3 represents a complex group in fish that has been shaped by gene duplication and diversification. Many putative transcription factor binding sites were identified in the predicted promoter region of Rbcasp3, including immune factor- and cancer signal-inducible sites. Rbcasp3, excluding the pro-domain, was expressed in Escherichia coli. The recombinant protein showed a detectable activity against the mammalian caspase 3/7-specific substrate DEVD-pNA, indicating a functional role in physiology. Quantitative real time PCR assay detected Rbcasp3 expression in all examined tissues, but with high abundance in blood, liver and brain. Transcriptional profiling of rock bream liver tissue revealed that challenge with lipopolysaccharides (LPS) caused prolonged up-regulation of Rbcasp3 mRNA whereas, Edwardsiella tarda (E. tarda) stimulated a late-phase significant transcriptional response. Rock bream iridovirus (RBIV) up-regulated Rbcasp3 transcription significantly at late-phase, however polyinosinic-polycytidylic acid (poly(I:C)) induced Rbcasp3 significantly at early-phase. Our findings suggest that Rbcasp3 functions as a cysteine-aspartate-specific protease and contributes to immune responses against bacterial and viral infections.

Original languageEnglish
Pages (from-to)99-110
Number of pages12
JournalFish and Shellfish Immunology
Volume33
Issue number1
DOIs
Publication statusPublished - 2012 Jul 1

Fingerprint

bream
caspase-3
Caspase 3
genomics
amino acid
rocks
Rocks
Amino Acids
Liver
rock
protein
functional role
Binding Sites
apoptosis
Tissue
immune response
Edwardsiella tarda
polyinosinic-polycytidylic acid
Poly I-C
liver

Keywords

  • Caspase 3
  • Hydrolyzing activity
  • Recombinant protein
  • Rock bream
  • Transcriptional analysis

ASJC Scopus subject areas

  • Aquatic Science
  • Environmental Chemistry

Cite this

Caspase 3 from rock bream (Oplegnathus fasciatus) : Genomic characterization and transcriptional profiling upon bacterial and viral inductions. / Elvitigala, Don Anushka Sandaruwan; Whang, Ilson; Premachandra, H. K A; Umasuthan, Navaneethaiyer; Oh, Myung Joo; Jung, Sung Ju; Yeo, Sang Yeob; Lim, Bong Soo; Lee, Jeong Ho; Park, Hae Chul; Lee, Jehee.

In: Fish and Shellfish Immunology, Vol. 33, No. 1, 01.07.2012, p. 99-110.

Research output: Contribution to journalArticle

Elvitigala, DAS, Whang, I, Premachandra, HKA, Umasuthan, N, Oh, MJ, Jung, SJ, Yeo, SY, Lim, BS, Lee, JH, Park, HC & Lee, J 2012, 'Caspase 3 from rock bream (Oplegnathus fasciatus): Genomic characterization and transcriptional profiling upon bacterial and viral inductions', Fish and Shellfish Immunology, vol. 33, no. 1, pp. 99-110. https://doi.org/10.1016/j.fsi.2012.04.008
Elvitigala, Don Anushka Sandaruwan ; Whang, Ilson ; Premachandra, H. K A ; Umasuthan, Navaneethaiyer ; Oh, Myung Joo ; Jung, Sung Ju ; Yeo, Sang Yeob ; Lim, Bong Soo ; Lee, Jeong Ho ; Park, Hae Chul ; Lee, Jehee. / Caspase 3 from rock bream (Oplegnathus fasciatus) : Genomic characterization and transcriptional profiling upon bacterial and viral inductions. In: Fish and Shellfish Immunology. 2012 ; Vol. 33, No. 1. pp. 99-110.
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