Molecular cloning of sheep T cell receptor γ and δ chain constant regions: unusual primary structure of γ chain hinge segments

Abstract

The primary structure of sheep T cell receptor (TcR) gamma and delta chain constant (C) regions has been determined by cDNA cloning. A comparison of the nucleotide and deduced amino acid sequences of the sheep chains with known human and mouse sequences shows that the primary structure of the immunoglobulin, transmembrane and cytoplasmic C gamma domains and all of the C delta region has been substantially conserved. However, the hinge or connector region of sheep gamma chains differs significantly from all known TcR chains. Clones representing two different sheep C gamma genes were isolated and both contain additional sequence in this region, making them the longest TcR chains so far identified. The hinge region of both sheep C gamma sequences contains two additional cysteine residues and a motif of five amino acids (TTESP or TTEPP) which has been triplicated in one of the clones. Other repetitive segments of 13-17 amino acids could also be identified suggesting that, as in the human C gamma 2 gene, this region of the sheep genes could have arisen from an exon duplication or triplication event. Southern blot analysis of sheep DNA confirmed the presence of one C delta gene and at least two C gamma genes. A restriction fragment length polymorphism that is probably associated with allelic sequence variation in the sheep C delta gene was detected in DNA from different animals. Although the essential structure of the gamma/delta TcR appears well conserved through evolution, the marked heterogeneity evident in the hinge region of gamma chains both within and between species, and particularly the presence of additional cysteine residues in the sheep sequences, may be of structural and functional importance.