Glycobiology Advance Access originally published online on March 16, 2007
Glycobiology 2007 17(6):586-599; doi:10.1093/glycob/cwm023
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Molecular cloning and characterization of the Caenorhabditis elegans 
1,3-fucosyltransferase family
3 Department of Biochemistry and Molecular Biology
4 Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center
5 Program in Molecular, Cell, and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104
6 Department of Molecular Cell Biology, Glycoimmunology Group, VU University Medical Center, Van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
1 To whom correspondence should be addressed: Tel: +1 (404) 727-5962; Fax: +1 (404) 727-2738; E-mail: rdcummi{at}emory.edu
Received on October 3, 2006; revised on February 21, 2007; accepted on February 22, 2007
The genome of Caenorhabditis elegans encodes five genes with homology to known 
1,3 fucosyltransferases (1,3FTs), but their expression and functions are poorly understood. Here we report the molecular cloning and characterization of these C. elegans 1,3FTs (CEFT-1 through -5). The open-reading frame for each enzyme predicts a type II transmembrane protein and multiple potential N-glycosylation sites. We prepared recombinant epitope-tagged forms of each CEFT and found that they had unusual acceptor specificity, cation requirements, and temperature sensitivity. CEFT-1 acted on the N-glycan pentasaccharide core acceptor to generate Man1-3(Man1-6)Manß1-4GlcNAcß1-4(Fuc1-3)GlcNAcß1-Asn. In contrast, CEFT-2 did not act on the pentasaccharide acceptor, but instead utilized a LacdiNAc acceptor to generate GalNAcß1-4(Fuc1-3)GlcNAcß1-3Galß1-4Glc, which is a novel activity. CEFT-3 utilized a LacNAc acceptor to generate Galß1-4(Fuc1-3)GlcNAcß1-3Galß1-4Glc without requiring cations. CEFT-4 was similar to CEFT-3, but its activity was enhanced by some divalent cations. Recombinant CEFT-5 was well expressed, but did not act on available acceptors. Each CEFT was optimally active at room temperature and rapidly lost activity at 37 °C. Promoter analysis showed that CEFT-1 is expressed in C. elegans eggs and adults, but its expression was restricted to a few neuronal cells at the head and tail. We prepared deletion mutants for each enzyme for phenotypic analysis. While loss of CEFT-1 correlated with loss of pentasaccharide core activity and core 1,3-fucosylated glycans in worms, loss of other enzymes did not correlate with any phenotypic changes. These results suggest that each of the 1,3FTs in C. elegans has unique specificity and expression patterns.
Key words: C. elegans / fucosyltransferase / cloning / N-glycans / neuronal cells
2 Present address: Department of Biochemistry, Emory University School of Medicine, 1510 Clifton Rd NE Suite #4001, Atlanta, GA 30322
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