Molecular cloning of a human fucosyltransferase gene that determines expression of the Lewis x and VIM-2 epitopes but not ELAM-1-dependent cell adhesion.
We have used the human Lewis blood group fucosyltransferase cDNA and cross-hybridization procedures to isolate a human gene that encodes a distinct fucosyltransferase. Its DNA sequence predicts a type II transmembrane protein whose sequence is identical to 133 of 231 amino acids at corresponding positions within the catalytic domain of the Lewis fucosyltransferase. When expressed by transfection in cultured cell lines, this gene determines expression of a fucosyltransferase capable of efficiently utilizing N-acetyllactosamine to form the Lewis x determinant (Gal beta 1----4[Fuc alpha 1----3]GlcNAc). By contrast, biochemical and flow cytometry analyses suggest that the enzyme cannot efficiently utilize the type II acceptor NeuNAc alpha 2----3Gal beta 1----4GlcNAc, to form the sialyl Lewis x determinant. In Chinese hamster ovary cells, however, the enzyme can determine expression of the alpha 2----3-sialylated, alpha 1----3-fucosylated structure known as VIM-2, a putative oligosaccharide ligand for ELAM-1. Cell adhesion assays using VIM-2-positive, sialyl Lewis x-negative transfected Chinese hamster ovary cells indicate that surface expression of the VIM-2 determinant is not sufficient to confer ELAM-1-dependent adhesive properties upon the cells. These results demonstrate that substantial structural similarities can exist between mammalian glycosyltransferases with closely related enzymatic properties, thus facilitating isolation of their cognate genes by cross-hybridization methods. The results further suggest that cell surface expression of the VIM-2 determinant is not necessarily sufficient to mediate ELAM-1-dependent cell adhesion.
Lowe
J B
JB
Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor 48109-0650.
Kukowska-Latallo
J F
JF
Nair
R P
RP
Larsen
R D
RD
Marks
R M
RM
Macher
B A
BA
Kelly
R J
RJ
Ernst
L K
LK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA; D019040:E-Selectin; D000939:Epitopes / Q000096:biosynthesis* / Q000276:immunology; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis / Q000276:immunology*; D016366:Open Reading Frames; D015183:Restriction Mapping
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D000939:Epitopes; D009844:Oligosaccharides; C070946:VIM-2 determinant; D004247:DNA; D005647:Fucosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase