Description of an ectothermic TCR coreceptor, CD8 alpha, in rainbow trout.
We have cloned the first CD8 alpha gene from an ectothermic source using a degenerate primer for Ig superfamily V domains. Similar to homologues in higher vertebrates, the rainbow trout CD8 alpha gene encodes a 204-aa mature protein composed of two extracellular domains including an Ig superfamily V domain and hinge region. Differing from mammalian CD8 alpha V domains, lower vertebrate (trout and chicken) sequences do not contain the extra cysteine residue (C strand) involved in the abnormal intrachain disulfide bridging within the CD8 alpha V domain of mice and rats. The trout membrane proximal hinge region contains the two essential cysteine residues involved in CD8 dimerization (alpha alpha or alpha beta) and threonine, serine, and proline residues which may be involved in multiple O-linked glycosylation events. Although the transmembrane region is well conserved in all CD8 alpha sequences analyzed to date, the putative trout cytoplasmic region differs and, in fact, lacks the consensus p56lck motif common to other CD8 alpha sequences. We then determined that the trout CD8 alpha genomic structure is similar to that of humans (six exons) but differs from that of mice (five exons). Additionally, Northern blotting and RT-PCR demonstrate that trout CD8 alpha is expressed at high levels within the thymus and at weaker levels in the spleen, kidney, intestine, and peripheral blood leukocytes. Finally, we show that trout CD8 alpha can be expressed on the surface of cells via transfection. Together, our results demonstrate that the basic structure and expression of CD8 alpha has been maintained for more than 400 million years of evolution.
Hansen
J D
JD
Basel Institute for Immunology, Basel, Switzerland. hansen@bii.ch
Strassburger
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001833:Body Temperature Regulation / Q000276:immunology; D016827:CD8 Antigens / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003593:Cytoplasm / Q000737:chemistry; D018076:DNA, Complementary / Q000302:isolation & purification; D005091:Exons; D005110:Extracellular Space / Q000737:chemistry / Q000276:immunology; D052058:Hinge Exons; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D007438:Introns; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000276:immunology*; D010446:Peptide Fragments / Q000737:chemistry; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012313:RNA / Q000096:biosynthesis; D051381:Rats; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry*
D016827:CD8 Antigens; D018076:DNA, Complementary; D007135:Immunoglobulin Variable Region; D008565:Membrane Proteins; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D012313:RNA