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Stable transfection of human β‐1,4N‐acetylgalactosaminyltransferase and α‐2,8‐sialyltransferase cDNAs in C6 rat glioma cells induces modifications in ganglioside metabolism
Author(s) -
Sottocornola Elena,
Colombo Irma,
Berra Bruno
Publication year - 2000
Publication title -
european journal of lipid science and technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.614
H-Index - 94
eISSN - 1438-9312
pISSN - 1438-7697
DOI - 10.1002/1438-9312(200011)102:11<673::aid-ejlt673>3.0.co;2-z
Subject(s) - glycosyltransferase , sialyltransferase , ganglioside , transfection , complementary dna , biochemistry , cell culture , biosynthesis , enzyme , biology , microbiology and biotechnology , chemistry , gene , genetics
Ganglioside distribution in cells undergoes deep modifications during physiological and pathological events, possibly depending on the activity of glycosyltransferases involved in their biosynthesis. To understand how the ganglioside pattern can be altered by the selective expression of specific glycosyltransferases, C6 rat glioma cell line was stably transfected with two human glycosyltransferase cDNAs: β ‐1,4N‐acetylgalactosaminyltransferase (GalNAcT) and α ‐2,8‐sialyltransferase (ST‐II). GalNAcT and ST‐II are key enzymes in ganglioside biosynthesis; whereas ST‐II synthesizes GD 3 , precursor of the “b” pathway, GalNAcT produces GM 2 , GD 2 and asialo‐GM 2 and it is, therefore, involved in “a”, “b” and “asialo” pathways. C6 cells were subjected to three independent transfections: one with a construct containing GalNAcT cDNA, one with a construct containing the ST‐II cDNA, and one with both constructs simultaneously. Whereas control cells present mainly N‐acetyl‐ and N‐glycolyl‐GM 3 , selected transfected clones show more complex ganglioside profiles: GalNAcT‐expressing cells are enriched in the “a” series gangliosides, ST‐II‐expressing cells synthesize the “b” series species, cells expressing contemporarily the two glycosyltransferases produce gangliosides of both series. Furthermore, among the selected clones, expression of GalNAcT and ST‐II correlates with changes in the ST‐I and ST‐IV activities, indicating that the switching on of the biosynthetic enzymes we investigated influences the activity of endogenous glycosyltransferases, possibly through the modification of the amount of their substrates or products.