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Molecular cloning and expression analysis of interferon-inducible transmembrane protein 1 in large yellow croaker
1Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, PR China.
Insights
Researchers identified a novel interferon-inducible transmembrane protein 1 (IFITM1) in large yellow croaker fish. This LycIFITM1 gene is expressed in various tissues and significantly upregulated by poly(I:C), suggesting a role in fish immune responses.
Area of Science:
- Immunology
- Molecular Biology
- Fish Biology
Background:
- Interferon-inducible transmembrane protein 1 (IFITM1) is crucial in human cells for cell adhesion and antiproliferative signaling.
- Understanding IFITM1 homologs in fish can elucidate conserved immune functions and evolutionary pathways.
Purpose of the Study:
- To clone and characterize the IFITM1 homologue from large yellow croaker (Pseudosciaena crocea).
- To investigate the expression patterns and immune inducibility of the LycIFITM1 gene.
Main Methods:
- cDNA cloning and sequencing of LycIFITM1.
- Bioinformatic analysis for protein structure and phylogenetic relationships.
- Quantitative real-time PCR to analyze gene expression in various tissues and upon poly(I:C) stimulation.
Main Results:
- The LycIFITM1 gene encodes a 124-amino acid protein with typical IFITM structural features, including transmembrane domains and a PKC phosphorylation site.
- Phylogenetic analysis places LycIFITM1 closer to fish IFITMs than mammalian counterparts.
- LycIFITM1 is constitutively expressed in multiple tissues and significantly upregulated in gills, kidney, heart, and spleen following poly(I:C) induction, peaking at 24 hours.
Conclusions:
- LycIFITM1 represents a novel fish member of the IFITM family.
- The inducible expression of LycIFITM1 suggests its involvement in the innate immune response of large yellow croaker, particularly against viral or microbial challenges.
- This finding contributes to understanding the evolution and function of IFITMs in aquatic vertebrates.
Abstract:
In human cells, interferon-inducible transmembrane protein 1 (IFITM1) is a component of protein complexes involved in homotypic adhesion and the transduction of antiproliferative signals. Here, we reported the cloning of an IFITM1 homologue from the spleen of large yellow croaker Pseudosciaena crocea (LycIFITM1). The complete cDNA of LycIFITM1 is 734 nucleotides (nt) encoding a protein of 124 amino acids (aa), with a putative molecular weight of 13.6 kDa. The deduced LycIFITM1 protein is significantly homologous to interferon-inducible transmembrane proteins (IFITMs) in mammals and fish, and has the typical structural features of IFITMs, including two transmembrane domains (residues 43-63 and 90-112, respectively) and one intracellular domain between them (residues 64-89), as well as one conserved protein kinase C (PKC) phosphorylation site (residues 65-67, SIK). Phylogenetic analysis showed that LycIFITM1 formed a cluster with fish IFITM, reflecting a relative distant evolutionary relationship from mammals. LycIFITM1 gene was constitutively expressed in various tissues examined, such as gills, intestine, liver, kidney, heart, spleen, muscle and blood. Upon induction with poly(I:C), LycIFITM1 gene expression was obviously up-regulated in gills, kidney, heart and spleen at 24h after stimulation, suggesting that LycIFITM1 may be involved in the immune response induced by poly(I:C). Time course analysis using real-time PCR showed that the mRNA levels of LycIFITM1 in spleen and kidney were quickly up-regulated by poly(I:C) and reached the peak at 24h post-induction (48.7- and 280.4-fold mRNA increases in spleen and kidney, respectively). The results suggest that the IFITM1 homologue from large yellow croaker may represent a novel member of IFITMs family in fish.
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