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

O Pleguezuelos, J Zou, C Cunningham, C J Secombes

Research output: Contribution to journalArticle

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Abstract

The full-length sequence of a second IL-1 beta gene (IL-1 beta 2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1 beta gene, IL-1 beta 2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 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, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation 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 beta. Comparison of the two trout genes reveals that the IL-1 beta 2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As 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) that 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 leukocytes by stimulation with lipopolysaccharide.

Original languageEnglish
Pages (from-to)1002-1011
Number of pages10
JournalImmunogenetics
Volume51
Publication statusPublished - 2000

Keywords

  • interleukin-1 beta
  • cDNA
  • cytokine
  • Oncorhynchus mykiss
  • gene organization
  • MESSENGER-RNA
  • INTERLEUKIN-1-BETA
  • PROTEIN
  • ALIGNMENT
  • CYTOKINE
  • MAMMALS
  • SEARCH
  • CELLS
  • ACID

Cite this

Cloning, sequencing, and analysis of expression of a second IL-1 beta gene in rainbow trout (Oncorhynchus mykiss). / Pleguezuelos, O ; Zou, J ; Cunningham, C ; Secombes, C J .

In: Immunogenetics, Vol. 51, 2000, p. 1002-1011.

Research output: Contribution to journalArticle

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abstract = "The full-length sequence of a second IL-1 beta gene (IL-1 beta 2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1 beta gene, IL-1 beta 2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 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, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation 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 beta. Comparison of the two trout genes reveals that the IL-1 beta 2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As 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) that 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 leukocytes by stimulation with lipopolysaccharide.",
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TY - JOUR

T1 - Cloning, sequencing, and analysis of expression of a second IL-1 beta gene in rainbow trout (Oncorhynchus mykiss)

AU - Pleguezuelos, O

AU - Zou, J

AU - Cunningham, C

AU - Secombes, C J

PY - 2000

Y1 - 2000

N2 - The full-length sequence of a second IL-1 beta gene (IL-1 beta 2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1 beta gene, IL-1 beta 2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 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, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation 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 beta. Comparison of the two trout genes reveals that the IL-1 beta 2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As 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) that 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 leukocytes by stimulation with lipopolysaccharide.

AB - The full-length sequence of a second IL-1 beta gene (IL-1 beta 2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1 beta gene, IL-1 beta 2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 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, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation 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 beta. Comparison of the two trout genes reveals that the IL-1 beta 2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As 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) that 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 leukocytes by stimulation with lipopolysaccharide.

KW - interleukin-1 beta

KW - cDNA

KW - cytokine

KW - Oncorhynchus mykiss

KW - gene organization

KW - MESSENGER-RNA

KW - INTERLEUKIN-1-BETA

KW - PROTEIN

KW - ALIGNMENT

KW - CYTOKINE

KW - MAMMALS

KW - SEARCH

KW - CELLS

KW - ACID

M3 - Article

VL - 51

SP - 1002

EP - 1011

JO - Immunogenetics

JF - Immunogenetics

SN - 0093-7711

ER -