Cloning of a rat alpha1,3-fucosyltransferase gene: a member of the fucosyltransferase IV family.
We report the cloning of a rat alpha1,3-fucosyltransferase gene (rFuc-T), isolated from a rat genomic library by a PCR-cross-hybridization based cloning approach using primers derived from the conserved region of human alpha1,3-Fuc-T sequences. Comparison of the rFuc-T predicted amino acid sequence with those of previously cloned human and murine fucosyltransferases showed highest degree of homology to murine Fuc-TIV (87% identity) and human Fuc-TIV (78% identity), with lower homology (41-49% identity) to Fuc-TIII, V, VI, and VII. COS-1 cells transfected with the rFuc-Tgene expressed a fucosyltransferase activity with type 2 (Gal beta1-->4GlcNAc)-containing oligosaccharides and the glycolipid acceptor neolactotetraosylceramide but only low activity with sialylated substrates; the SSEA-1/Le(x) antigen was detected in transfected cells by immunocytochemistry. Based on these results, we surmise that rFuc-T is a member of the fucosyltransferase IV family. Northern blot analysis with a rFuc-T specific probe indicated a major transcript of 4.2 kb most abundantly expressed in rat spleen; minor transcripts of different sizes were detected in several tissues, including rat brain.
Sajdel-Sulkowska
E M
EM
Department of Biomedical Sciences, E.K. Shriver Center, Waltham, MA 02254, USA.
Smith
F I
FI
Wiederschain
G
G
McCluer
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology; D019556:COS Cells / Q000378:metabolism; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D016256:Lewis X Antigen / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014018:Tissue Distribution; D014162:Transfection
D016256:Lewis X Antigen; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase