Cloning, sequencing, and analysis of expression of a second IL-1 beta genein rainbow trout (Oncorhynchus mykiss)

The full-length sequence of a second IL-1 beta gene (IL-1 beta 2) in rainbo w trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1 bet a gene, IL-1 beta 2 is organized into six exons/five introns. There are onl y small differences in their intron/exon sizes, with the exception of intro n 3, which is 334 bp smaller in IL-1 beta 2. The transcript encoded by the IL-1 beta 2 gene contains a 5' untranslated region (UTR) of 121 bp, followe d by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains sev en instability attta motifs, typical of inflammatory genes, and a polyadeny lation site II bp upstream of a 17-bp poly(A) tail. The predicted 254 amino acid sequence of the second IL-1 beta gene has 82% similarity to the first gene, 45% similarity to carp IL-1 beta, and 40% similarity to human IL-1 b eta. Comparison of the two trout genes reveals that the IL-1 beta 2 gene ha s a deletion of 9 bases in exon 3 and an altered splicing site at the 5' en d of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. A s with other nonmammalian IL-1 beta genes, no interleukin-converting enzyme (ICE) cut site has been found but the alignment of the amino acid sequence with Other species shows a possible cut site between Arg(89) and Ala(90) t hat would give arise to a 165-amino acid mature peptide. Expression studies performed by RT-PCR using primers specific for the IL-1 beta 2 transcript revealed a clear dose-dependent induction of this gene in cultured trout le ukocytes by stimulation with lipopolysaccharide.