Porcine alpha1,3-galactosyltransferase: tissue-specific and regulated expression of splicing isoforms.
Expression of the Gal alpha1,3 Gal epitope on membrane glycolipids and glycoproteins is known to vary widely from one tissue to another. In the course of studying the mechanisms underlying this variability, we have isolated from pig cDNA four sequences corresponding to four isoforms of alpha1,3-galactosyltransferase (alpha1,3GT), the Golgi enzyme that links galactose in alpha1,3 on the galactose residue of N-acetyllactosamine. The isoforms differ from each other in the alternative presence of two nucleotide stretches of 36 and 63 base pairs in a segment encoding the stem region of the protein. Stable expression experiments show that all four isoenzymes can confer alpha-galactosyltransferase activity to HeLa cells, and that they are all located within the Golgi compartment, indicating that variations in length in the stem region do not affect enzyme activity or cellular localization. Analysis of RNA from different pig organs and cells shows quantitative differences between tissues in levels of alpha1,3GT, as well as qualitative differences, the four isoforms being unequally represented in different tissues.
Vanhove
B
B
Institut National de la Santé et de la Recherche Médicale U437, Unité de Recherche sur l'Immunointervention dans les Allo et Xénotransplantations, Nantes, France. bvanhove@sante.univ-nantes.fr
Goret
F
F
Soulillou
J P
JP
Pourcel
C
C
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004730:Endothelium, Vascular / Q000201:enzymology; D005700:Galactosyltransferases / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000201:enzymology*; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010957:Plasmids; D012313:RNA / Q000032:analysis; D016415:Sequence Alignment; D013552:Swine
D018076:DNA, Complementary; D007527:Isoenzymes; D012313:RNA; D005700:Galactosyltransferases