TY - JOUR
T1 - Activation of two new α(1,3)fucosyltransferase activities in Chinese hamster ovary cells by 5-azacytidine
AU - Potvin, Barry
AU - Stanley, Pamela
PY - 1991/12
Y1 - 1991/12
N2 - Several mammalian α(1,3)fucosyltransferases (α[1,3]Fuc-T) that synthesize carbohydrates containing α(1,3)fucosylated lactosamine units have been identified. Although Chinese hamster ovary (CHO) cells do not express α(1,3)Fuc-T activity, the rare mutants LEC11 and LEC12, isolated after mutagenesis or DNA transfection, each express an α(1,3)Fuc-T that may be distinguished by several criteria. Two new CHO mutants possessing α(1,3)Fuc-T activity (LEC29 and LEC30) have now been isolated after treatment of a CHO cell population with 5-azacytidine (5-AzaC), ethylnitrosourea (ENU), or 5-AzaC followed by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Like LEC12, both mutants possess an N-ethylmaleimide-resistant α(1,3)Fuc-T activity that can utilize a variety of acceptors and both express the Lewis X (Lex) determinant {Galβ[1,4](Fucα[1,3])GlcNAcβ1} but not the sialylα(2,3)Lex determinant on cell-surface carbohydrates. However, LEC29 and LEC30 may be distinguished from LEC11 and LEC12, as well as from each other, on the basis of their unique patterns of lectin resistance and their abilities to bind the VIM-2 monoclonal antibody that recognizes carbohydrates terminating in NeuNAcα(2,3)Galβ(1,4)GlcNAcβ(1,3)Galβ(1,4)(Fucα[1,3]) GlcNAcβ and also by the different in vitro substrate specificities and kinetic properties of their respective α(1,3)Fuc-T activities. The combined data provide good evidence that the LEC29 and LEC30 α(1,3)Fuc-Ts are novel transferases encoded by distinct gene products.
AB - Several mammalian α(1,3)fucosyltransferases (α[1,3]Fuc-T) that synthesize carbohydrates containing α(1,3)fucosylated lactosamine units have been identified. Although Chinese hamster ovary (CHO) cells do not express α(1,3)Fuc-T activity, the rare mutants LEC11 and LEC12, isolated after mutagenesis or DNA transfection, each express an α(1,3)Fuc-T that may be distinguished by several criteria. Two new CHO mutants possessing α(1,3)Fuc-T activity (LEC29 and LEC30) have now been isolated after treatment of a CHO cell population with 5-azacytidine (5-AzaC), ethylnitrosourea (ENU), or 5-AzaC followed by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Like LEC12, both mutants possess an N-ethylmaleimide-resistant α(1,3)Fuc-T activity that can utilize a variety of acceptors and both express the Lewis X (Lex) determinant {Galβ[1,4](Fucα[1,3])GlcNAcβ1} but not the sialylα(2,3)Lex determinant on cell-surface carbohydrates. However, LEC29 and LEC30 may be distinguished from LEC11 and LEC12, as well as from each other, on the basis of their unique patterns of lectin resistance and their abilities to bind the VIM-2 monoclonal antibody that recognizes carbohydrates terminating in NeuNAcα(2,3)Galβ(1,4)GlcNAcβ(1,3)Galβ(1,4)(Fucα[1,3]) GlcNAcβ and also by the different in vitro substrate specificities and kinetic properties of their respective α(1,3)Fuc-T activities. The combined data provide good evidence that the LEC29 and LEC30 α(1,3)Fuc-Ts are novel transferases encoded by distinct gene products.
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U2 - 10.1091/mbc.2.12.989
DO - 10.1091/mbc.2.12.989
M3 - Article
C2 - 1724918
AN - SCOPUS:0026264826
SN - 1059-1524
VL - 2
SP - 989
EP - 1000
JO - Molecular biology of the cell
JF - Molecular biology of the cell
IS - 12
ER -