Cloning of a novel rainbow trout (Oncorhynchus mykiss) CC chemokine with a fractalkine-like stalk and a TNF decoy receptor using cDNA fragments containing AU-rich elements.
An activation-specific cDNA library was made from phytohaemagglutinin (PHA)-activated haematopoietic cells of the rainbow trout (Oncorhynchus mykiss) using the technique of suppression subtractive hybridization. Several immune system genes were identified, including an interleukin (IL)1 receptor related protein and two invariant chain-like proteins. Many clones showed no similarity by BLAST search, but had AU-rich elements. These fragments were labelled and used for hybridization with a PHA-activated head kidney cDNA library. Several immune system genes were isolated by this technique, including a tumour necrosis factor (TNF) decoy receptor and a novel chemokine, designated trout chemokine 2. The TNF receptor is 285 amino acids in length and is 32-36% identical to a brook trout and human homologue. The CC chemokine is 44% identical at the amino acid level to a carp CC chemokine and approximately 20% identical to several mammalian CC chemokines. However, it has a 91 amino acid stalk-like structure at its COOH end, which is similar to the glycosylated stalk of fractalkine, a mammalian CX(3)C chemokine. In summary, AU-rich fragments obtained from an activation-specific library proved useful as hybridization probes for isolating trout immune system genes.
Liu
Lei
L
Department of Immunology and Microbiology, Wayne State University School of Medicine, 540 East Canfield Avenue, Detroit, MI 48201, USA.
Fujiki
Kazuhiro
K
Dixon
Brian
B
Sundick
Roy S
RS
D000225:Adenine; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D054428:Chemokine CX3CL1; D019742:Chemokines, CC / Q000235:genetics*; D020523:Chemokines, CX3C / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D029941:Fish Proteins / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics*; D010835:Phytohemagglutinins / Q000494:pharmacology; D021382:Protein Sorting Signals; D012313:RNA; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014529:Uridine
C515538:CX3CL1 protein, human; D054428:Chemokine CX3CL1; D019742:Chemokines, CC; D020523:Chemokines, CX3C; D018076:DNA, Complementary; D029941:Fish Proteins; D008565:Membrane Proteins; D010835:Phytohemagglutinins; D021382:Protein Sorting Signals; D018124:Receptors, Tumor Necrosis Factor; D012313:RNA; D000225:Adenine; D014529:Uridine