Molecular cloning and expression of a bovine alpha(1,3)-fucosyltransferase gene homologous to a putative ancestor gene of the human FUT3-FUT5-FUT6 cluster

Abstract

Only one bovine gene, corresponding to the human cluster of genes FUT3-FUT5-FUT6, was found by Southern blot analysis. The cognate bovine alpha(1,3)-fucosyltransferase shares 67.3, 69.0, and 69.3% amino acid sequence identities with human FUC-T3, FUC-T5, and FUC-T6 enzymes, respectively. As revealed by protein sequence alignment, potential sites for asparagine-linked glycosylation and conserved cysteines, the bovine enzyme is an intermediate between FUC-T3, FUC-T5, and FUC-T6 human enzymes. Transfected into COS-7 cells, the bovine gene induced the synthesis of an alpha(1, 3)-fucosyltransferase enzyme with type 2 substrate acceptor pattern specificity and induced expression of fucosylated type 2 epitopes (Lex and sialyl-Lex), but not of type 1 structures (Lea or sialyl-Lea), suggesting that it has an acceptor specificity similar to the human plasma FUC-T6. However, no enzyme activity was detected in bovine plasma. Gene transcripts are detected on tissues such as bovine liver, kidney, lung, and brain. The type 2 sialyl-Lex epitope was found in renal macula densa and biliary ducts, and Lex and Ley epitopes were detected on the brush border of epithelial cells of small and large intestine, suggesting a tissue distribution closer to human FUC-T3, but fucosylated type 1 structures (Lea, Leb, or sialyl-Lea) were not detected at all in any bovine tissue. Analysis of genetic distances on a combined phylogenetic tree of fucosyltransferase genes suggests that the bovine gene is the orthologous homologue of the ancestor of human genes constituting the present FUT3-FUT5-FUT6 cluster.