Molecular cloning, expression and functional characterization of a teleostan cytokine-induced apoptosis inhibitor from rock bream (Oplegnathus fasciatus).
Apoptosis plays a key role in the physiology of multicellular organisms and is regulated by different promoting and inhibitory mechanisms. Cytokine-induced apoptotic inhibitor (CIAPI) was recently identified as a key factor involved in apoptosis inhibition in higher vertebrate lineages. However, most of the CIAPIs of lower vertebrate species are yet to be characterized. Herein, we molecularly characterized a teleostan counterpart of CIAPI from rock bream (Oplegnathus fasciatus), designating as RbCIAPI. The complete coding region of RbCIAPI was consisted of 942 nucleotides encoding a protein of 313 amino acids with a predicted molecular mass of ~33 kDa. RbCIAPI gene exhibited a multi-exonic architecture, consisting 9 exons interrupted by 8 introns. Protein sequence analysis revealed that RbCIAPI shares significant homology with known CIAPI counterparts, and phylogenetic reconstruction confirmed its closer evolutionary relationship with its fish counterparts. Ubiquitous spatial distribution of RbCIAPI was detected in our quantitative real time polymerase chain reaction (qPCR) analysis, where more prominent expression levels were observed in the blood and liver tissues. Moreover, the RbCIAPI basal transcription level was found to be modulated by different bacterial and viral stimuli, which could be plausibly supported by our previous observations on the transcriptional modulation of the caspase 3 counterpart of rock bream (Rbcasp3) in response to the same stimuli. In addition, our in vitro functional assay demonstrated that recombinant RbCIAPI could detectably inhibit the proteolysis activity of recombinant Rbcasp3. Collectively, our preliminary results suggest that RbCIAPI may play an anti-apoptotic role in rock bream physiology, likely by inhibiting the caspase-dependent apoptosis pathway. Therefore, RbCIAPI potentially plays an important role in host immunity by regulating the apoptosis process under pathogenic stress.
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Development Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
Don Anushka Sandaruwan
DA
Elvitigala
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
H K A
HK
Premachandra
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Development Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea. Electronic address: ilsonwhang@hanmail.net.
Ilson
I
Whang
Department of Biotechnology, Division of Applied Chemistry & Biotechnology, Hanbat National University, Daejeon 305-719, Republic of Korea.
Sang-Yeob
SY
Yeo
Division of Marine Environment & Bioscience, Korea Maritime University, Busan 606-791, Republic of Korea.
Cheol Young
CY
Choi
Genetics & Breeding Research Center, National Fisheries Research & Development Institute, Geoje 656-842, Republic of Korea.
Jae Koo
JK
Noh
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Development Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea. Electronic address: jehee@jejunu.ac.kr.
Jehee
J
Lee
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001798:Blood Proteins / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D054357:Chemokine CXCL10 / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003531:Cypriniformes / Q000235:genetics / Q000276:immunology*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000502:physiology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D059467:Transcriptome
D001798:Blood Proteins; C489114:CIAPIN1 protein, human; D054357:Chemokine CXCL10; D029941:Fish Proteins; D047908:Intracellular Signaling Peptides and Proteins; D053148:Caspase 3
Caspase inhibitory activity; Cytokine-induced apoptosis inhibitor; Genomic gene structure; Rock bream; Transcriptional analysis