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MOLECULAR CHARACTERIZATION AND TRANSCRIPTIONAL ASPECTS WITH IMMUNE RESPONSIVE mRNA EXPRESSION PROFILES OF TWO GENES, MAPK1 AND INTERFERON INDUCED 44 LIKE (IFI44) COUNTERPART IDENTIFIED FROM DISK ABALONE (Haliotis discus discus)

N. C. N. Perera1,2*, G. I. Godahewa and Jehee Lee
E-mail: chathunp@gmail.com
 
1Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea
2Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea

Abalone immune responses cause by the induction of two different signal transduction pathways, the NF-κB and Mitogen-activated protein kinase (MAPK). Different MAPKs identified in all eukaryotic cells regulate diverse cellular functions for instances, gene expression, motility, metabolism, cell survival, proliferation and differentiation. Interferons (IFNs) are cytokines which released by the host as the response for the presence of pathogen, that mainly characterized by antiviral and immunomodulatory activities. Though, IFN has not been publicized in lower order invertebrate species few studies have conducted on interferon induced proteins in abalone. To date previous studies has proven that IFNs exert antiviral and immune-regulatory activities in different species and it can be assumed that the interferon induced protein activities are mediators of these pleiotropic effects. This study examined the characteristics at molecular and transcriptional level of the MAPK1 of disk abalone (AbMAPK1) and interferon induced 44 like counterpart of disc abalone (AbIFI44). AbMAPK1 encoded 1101 nucleotides along with 5' untranslated region (UTR) of 23 nucleotides and 3' UTR of 448 nucleotides with a molecular weight of 42.2 kDa and a pi of 6.28 where AbIFI44 encoded 711 bp which could be coded 236 amino acids polypeptide with a 26 kDa molecular mass and 8.3 pI. Polyadenylation signals (1440AATAAA1445, 1457AATAAA1462) and poly-A tail were identified at the end of 3' UTR in AbIFI44. AbMAPK1 aa sequence discovered two conserved protein domains. Further, there are ATP binding sites, polypeptide binding sites and active sites that important for the function of the MAPK1 gene. AbIFI44 showed that presence of GTP/Mg2+ binding sites which might be collectively involved in transcription and signal transduction in abalone. Pairwise alignments revealed that AbMAPK1 shared 87.9% of the highest identity and 92.5% of similarity with Aplysia californica. AbIFI44 shared quit low highest identity (23.5%) and similarity (40.3%) with teleostean IFI44 counterpart (Takifugu rubripes). AbMAPK1 closely cladded with A. californica. gill tissue shows the highest expressional level of AbMAPK1 where AbIFI44 mRNA markedly expressed in hepatopancreas, digestive tract and gill. Kinetic expression of AbMAPK1 transcripts in haemocytes tissue showed an early response with the Vibrio parahaemolyticus (3 hr p.i.), VHS virus (12 hr p.i) and Listeria monocytogenes (6 hr p.i). Kinetic expression of AbIFI44 transcripts in gill tissue was examined upon viral hemorrhagic septicemia virus (VHSV) challenge and significant mRNA upregulation was noted at 12 h and 72 h post infection. Collectively, it could be suggested that AbMAPK1 and AbIFI44 contributes to the antibacterial and antiviral defense mechanisms in abalone.

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