Molecular characterisation of the IRAK1 gene and its expression analysis in the Indian catfish Clarias magur, following Aeromonas hydrophila infection challenge
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Abstract
Interleukin-1 receptor associated kinase 1 (IRAK1) is a crucial downstream signaling mediator of the Interleukin-1 (IL-1) and Toll-like receptor (TLR) signal transduction pathways, responsible for sensing invading pathogens or endogenous danger signals and initiating appropriate immune responses. In this study, the partial cDNA sequence of IRAK1 gene was amplified, cloned, characterised and its modulation in response to Aeromonas hydrophila infection was studied in the Indian catfish Clarias magur, a promising fish species for aquaculture in the Indian subcontinent. The magur IRAK1 (mIRAK1) cDNA sequence of 590 bp was obtained by cloning and was found to encode a putative protein of 196 amino acid residues. This protein contains a serine/threonine kinase catalytic domain (STKcD) with activation sites responsible for kinase activity having a calculated isoelectric point (pI) of 6.65 and molecular weight of 21.68 kDa. Phylogenetically, mIRAK1 clustered together well with other members of the catfish family and showed the highest identity with pangasius catfish. Presence of highly conserved elements, including a conserved STKcD domain and cycline-dependent kinase 1 (CDK1) motif, were revealed by multiple sequence alignments in mIRAK1. Basal expression analysis indicated that mIRAK1 was widely expressed in all examined tissues, with the lowest expression in muscle and the highest in liver. Following A. hydrophila infection challenge, mIRAK1 expression was significantly up-regulated in vital immunological organs such as kidney, spleen, intestine and liver at 3 to 24 h post-infection. These results emphasise the critical role of mIRAK1 in host defence mediated by TLR and IL-1 signalling pathways during A. hydrophila infection.
Keywords: Basal expression, cDNA sequence, Immune gene expression, TLR and IL-1 signalling pathways
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