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Published on: September 12, 2013
Identification and characterization of a fish natural resistance-associated macrophage protein (NRAMP) cDNA
J P Saeij1, G F Wiegertjes, R J Stet
1Cell Biology and Immunology Group, Wageningen Institute of Animal Sciences, Wageningen Agricultural University, P.O. Box 338, 6700 AH Wageningen, The Netherlands.
Abstract:
The mouse Lsh/Ity/Bcg locus regulates natural resistance to intracellular pathogens, and the Nramp1 gene was isolated as its candidate. Nramp is part of a small family of at least two genes, Nramp1 and Nramp2. In the present study, a full-length cDNA for carp NRAMP has been isolated and characterized. Nucleotide and predicted amino acid sequence analysis indicate that the carp NRAMP encodes a 548 amino acid membrane protein with 12 putative transmembrane domains, two N-linked glycosylation sites, and an evolutionarily conserved consensus transport motif. The peptide sequence identity among carp and human NRAMP2 is 78%, and 65% with human NRAMP1. Reverse transcription-polymerase chain reaction revealed that carp NRAMP is ubiquitously expressed. Phylogenetic analysis, using neighbor-joining, showed that the carp NRAMP protein clustered together with mammalian NRAMP2 proteins.
Insights
Researchers characterized carp NRAMP, a gene crucial for natural resistance to pathogens. This study identified its structure, ubiquitous expression, and evolutionary relationship to mammalian NRAMP2, advancing our understanding of immune system genes in fish.
Area of Science:
- Molecular Biology
- Immunology
- Comparative Genomics
Background:
- The Lsh/Ity/Bcg locus in mice is key for natural resistance against intracellular pathogens.
- The Nramp1 gene was identified as a candidate gene within this locus.
- The Nramp gene family includes at least Nramp1 and Nramp2.
Purpose of the Study:
- To isolate and characterize the full-length cDNA of carp NRAMP.
- To analyze the structural and evolutionary features of carp NRAMP.
- To determine the expression pattern of carp NRAMP.
Main Methods:
- cDNA isolation and sequencing.
- Bioinformatic analysis of nucleotide and amino acid sequences.
- Reverse transcription-polymerase chain reaction (RT-PCR) for expression analysis.
- Phylogenetic analysis using neighbor-joining.
Main Results:
- A full-length carp NRAMP cDNA was isolated, encoding a 548 amino acid membrane protein.
- The protein features 12 putative transmembrane domains, two N-linked glycosylation sites, and a conserved transport motif.
- Carp NRAMP shares 78% peptide sequence identity with human NRAMP2 and 65% with human NRAMP1.
- RT-PCR confirmed ubiquitous expression of carp NRAMP.
- Phylogenetic analysis placed carp NRAMP within the NRAMP2 cluster of mammalian proteins.
Conclusions:
- Carp NRAMP is a functional homolog of mammalian NRAMP2.
- The identified carp NRAMP protein possesses conserved structural and functional domains.
- Ubiquitous expression suggests a broad role for carp NRAMP in fish physiology.
