A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group alpha(1,3/1,4)fucosyltransferase.
The stage-specific embryonic antigen SSEA-1 is a cell-surface oligosaccharide molecule expressed with temporal precision during the murine preimplantation period and implicated in adhesive events involving the process of compaction. We used a mammalian transient expression system to isolate a cloned human cDNA that determines expression of the SSEA-1 molecule. The cDNA sequence predicts a type II transmembrane protein with a domain structure similar to mammalian glycosyltransferases, but without primary sequence similarity to these enzymes. The carboxy-terminal domain of this protein was shown to be catalytically active as a fucosyltransferase when expressed in COS-1 cells as a portion of a secreted protein A fusion peptide. The enzyme is an exceptional glycosyltransferase in that it can use both type I and type II oligosaccharides as acceptor substrates to generate subterminal Fuc alpha(1,4)- and Fuc alpha(1,3)-linkages, respectively, in a manner analogous to the human Lewis blood group fucosyltransferase. Southern blot analysis shows that the cDNA corresponds to sequences syntenic to the Lewis locus on chromosome 19. These results indicate that this cDNA is the product of the human Lewis blood group locus, provide genetic confirmation of the hypothesis that this enzyme can catalyze two distinct transglycosylation reactions, and outline an approach to the isolation of other sequences that determine expression of developmentally regulated oligosaccharide antigens.
Kukowska-Latallo
J F
JF
Howard Hughes Medical Institute, Ann Arbor, Michigan.
Larsen
R D
RD
Nair
R P
RP
Lowe
J B
JB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005647:Fucosyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000235:genetics*; D016256:Lewis X Antigen / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012689:Sequence Homology, Nucleic Acid
D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen; D008565:Membrane Proteins; D004247:DNA; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase