Molecular cloning and characterization of an alpha1,3 fucosyltransferase, CEFT-1, from Caenorhabditis elegans.
We report on the identification, molecular cloning, and characterization of an alpha1,3 fucosyltransferase (alpha1,3FT) expressed by the nematode, Caenorhabditis elegans . Although C. elegans glycoconjugates do not express the Lewis x antigen Galbeta1-->4[Fucalpha1-->3]GlcNAcbeta-->R, detergent extracts of adult C.elegans contain an alpha1,3FT that can fucosylate both nonsialylated and sialylated acceptor glycans to generate the Lexand sialyl Lexantigens, as well as the lacdiNAc-containing acceptor GalNAcbeta1-->4GlcNAcbeta1-->R to generate GalNAcbeta1-->4 [Fucalpha1-->3]GlcNAcbeta1-->R. A search of the C.elegans genome database revealed the existence of a gene with 20-23% overall identity to all five cloned human alpha1,3FTs. The putative cDNA for the C.elegans alpha1,3FT (CEFT-1) was amplified by PCR from a cDNA lambdaZAP library, cloned, and sequenced. COS7 cells transiently transfected with cDNA encoding CEFT-1 express the Lex, but not sLexantigen. The CEFT-1 in the transfected cell extracts can synthesize Lex, but not sialyl Lex, using exogenous acceptors. A second fucosyltransferase activity was detected in extracts of C. elegans that transfers Fuc in alpha1,2 linkage to Gal specifically on type-1 chains. The discovery of alpha-fucosyltransferases in C. elegans opens the possibility of using this well-characterized nematode as a model system for studying the role of fucosylated glycans in the development and survival of C.elegans and possibly other helminths.
DeBose-Boyd
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Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, BRC 417, 975 N.E. 10th Street, Oklahoma City, OK 73104, USA.
Nyame
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Cummings
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics*; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D017238:Genes, Helminth; D016256:Lewis X Antigen / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009844:Oligosaccharides / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D000080506:Sialyl Lewis X Antigen; D014162:Transfection
D018076:DNA, Complementary; D016256:Lewis X Antigen; D009844:Oligosaccharides; D011994:Recombinant Proteins; D000080506:Sialyl Lewis X Antigen; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase