Molecular Cloning and Characterization of CFT1, a Developmentally Regulated Avian α(1,3)-Fucosyltransferase Gene

• Published in: The Journal of biological chemistry 1996-12, Vol.271 (51), p.32960-32967
• Main Authors: Lee, Kelvin P., Carlson, Louise M., Woodcock, Juliana B., Ramachandra, Nandini, Schultz, Terrie L., Davis, Thomas A., Lowe, John B., Thompson, Craig B., Larsen, Robert D.
• Format: Article
• Language: English
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; Amino Acid Sequence; Animals; ANTIGENE; ANTIGENOS; ARN MENSAJERO; ARN MESSAGER; B-Lymphocytes - enzymology; Base Sequence; BOLSA DE FABRICIO; BOURSE DE FABRICIUS; Chickens - blood; Chickens - immunology; CLONACION; CLONAGE; Cloning, Molecular; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; COS Cells; DESARROLLO EMBRIONARIO; DEVELOPPEMENT EMBRYONNAIRE; EXPRESION GENICA; EXPRESSION DES GENES; Fucosyltransferases - genetics; GENE; Gene Expression Regulation, Developmental; GENES; GLANDE THYMUS; GLANDULA DEL TIMO; GLICOSILTRANSFERASAS; GLYCOSYLTRANSFERASE; Lewis X Antigen - genetics; Lewis X Antigen - metabolism; LINFOCITOS; LYMPHOCYTE; Membrane Glycoproteins - genetics; Molecular Sequence Data; POLLO; POULET; Restriction Mapping; RNA, Messenger - genetics; SECUENCIA NUCLEOTIDICA; Sequence Alignment; Sequence Homology, Amino Acid; SEQUENCE NUCLEOTIDIQUE; Tissue Distribution
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.271.51.32960

Although coordinate expression of carbohydrate epitopes during development is well described, mechanisms which regulate this expression remain largely unknown. In this study we demonstrate that developing chicken B cells express the LewisX terminal oligosaccharide structure in a stage-specific manner. To examine regulation of this expression, we have cloned and expressed the chicken α(1,3)-fucosyltransferase gene involved in LewisX biosynthesis, naming it chicken fucosyltransferase 1 (CFT1). CFT1 is characterized by a single long open reading frame of 356 amino acids encoding a type II transmembrane glycoprotein. The domain structure and predicted amino acid sequence are highly conserved between CFT1 and mammalian FucTIV genes (52.8% and 46.3% identity to mouse and human respectively). In vitro CFT1 fucosyltransferase activity utilizes LacNAc > 3′sialyl-LacNAc acceptors with almost no utilization of other neutral type II (lactose, 2-fucosyllactose), or type I (lacto-N-biose I) acceptors. CFT1-transfected cells make cell surface LewisX (COS-7) and LewisX+ VIM-2 structures (Chinese hamster ovary). CFT1 gene expression is tissue-specific and includes embryonic thymus and bursa. Furthermore, expression of the CFT1 gene and cell surface LewisX structures are closely linked during B cell development. These findings reveal the evolutionary conservation between nonmammalian and mammalian α(1,3)-fucosyltransferase genes and demonstrate a role for fucosyltransferase gene regulation in the developmental expression of oligosaccharide structures.