Abstract
The structures of the canine, rabbit, bovine and equine EIF2AK2 genes were determined. Each of these genes has a 5' non-coding exon as well as 15 coding exons. All of the canine, bovine and equine EIF2AK2 introns have consensus donor and acceptor splice sites. In the equine EIF2AK2 gene, a unique single nucleotide polymorphism that encoded a Tyr329Cys substitution was detected. Regulatory elements predicted in the promoter region were conserved in ungulates, primates, rodents, Afrotheria (elephant) and Insectifora (shrew). Western clawed frog and fugu EIF2AK2 gene sequences were detected in the USCS Genome Browser and compared to those of other vertebrate EIF2AK2 genes. A comparison of EIF2AK2 protein domains in vertebrates indicates that the kinase catalytic domains were evolutionarily more conserved than the nucleic acid-binding motifs. Nucleotide substitution rates were uniform among the vertebrate sequences with the exception of the zebrafish and goldfish EIF2AK2 genes, which showed substitution rates about 20% higher than those of other vertebrates. FISH was used to physically assign the horse and cattle genes to chromosome locations, ECA15q24–q25 and BTA11q12–15, respectively. Comparative mapping data confirmed conservation of synteny between ungulates, humans and rodents.
Highlights
Additional alternative splice variants that differ in their 3’ non-coding regions were previously listed in the Celera database; the HSAEIF2AK2 mRNA BM473760 contains exon 17 extended with a significant portion of un-spliced intron but exons and 19 are absent in this transcript, while the transcript AA639687, contains exons 17, 18 and 19 but not intron 17
We report here characterization of the structures of the equine (Equus caballus; ECA), canine (Canis familiaris; CFA) and rabbit (Oryctolagus cuniculus; OCU) EIF2AK2 genes
The equine and bovine EIF2AK2 genes were localized on horse and cattle chromosomes, respectively, by fluorescent in situ hybridization (FISH) with several bacterial artificial chromosome (BAC) clones
Summary
The eukaryotic translation initiation factor 2-alpha kinase 2 (EIF2AK2, known as PKR or PRKR) is an important component of the host innate immune. Activation by dsRNA causes autophosphorylation of EIF2AK2 and allows this kinase to phosphorylate its natural substrate, the 1-alpha subunit of eukaryotic translation initiation factor (EIF2S1, known as eIF-2alpha). Phosphorylation of this initiation factor results in inhibition of protein translation and viral replication [25]. Previous reports described the structures of the human and mouse genes encoding the translation initiation factor 2-alpha kinase 2. Three alternative splicing acceptors in the second HSAEIF2AK2 exon have been reported [12]. The equine and bovine EIF2AK2 genes were localized on horse and cattle chromosomes, respectively, by fluorescent in situ hybridization (FISH) with several bacterial artificial chromosome (BAC) clones
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