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Volume 271, Number 51, Issue of December 20, 1996 pp. 32960-32967
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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

(Received for publication, July 3, 1996, and in revised form, September 17, 1996)

Kelvin P. Lee Dagger , Louise M. Carlson , Juliana B. Woodcock Dagger , Nandini Ramachandra par , Terrie L. Schultz ** , Thomas A. Davis Dagger , John B. Lowe par Dagger Dagger , Craig B. Thompson §§ and Robert D. Larsen **

From the Dagger  Immune Cell Biology Program, Naval Medical Research Institute, Bethesda, Maryland 20889,  NIAID, National Institutes of Health, Bethesda, Maryland 20892, the par  Howard Hughes Medical Institute, University of Michigan, Ann Arbor, Michigan 48109, ** Glycomed Inc., Alameda, California 94501, the Dagger Dagger  Department of Pathology, University of Michigan, Ann Arbor, Michigan 48109, and the §§ Gwenn Knapp Center for Lupus and Immunology Research, Department of Medicine, Department of Molecular Genetics and Cell Biology, Howard Hughes Medical Institute, University of Chicago, Chicago, Illinois 60637

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 alpha (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 alpha (1,3)-fucosyltransferase genes and demonstrate a role for fucosyltransferase gene regulation in the developmental expression of oligosaccharide structures.


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