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J Biol Chem, Vol. 273, Issue 41, 26779-26789, October 9, 1998

Single Glycosyltransferase, Core 2 16-N-acetylglucosaminyltransferase, Regulates Cell Surface Sialyl-Le^x Expression Level in Human Pre-B Lymphocytic Leukemia Cell Line KM3 Treated with Phorbolester

Mitsuru Nakamura, Takashi Kudo^¶, Hisashi Narimatsu^¶, Yusuke Furukawa, Jiro Kikuchi, Shinji Asakura^**, Wei Yang^**, Satsuki Iwase, Kiyohiko Hatake^§§, and Yasusada Miura^§§

From the  Division of Hemopoiesis, and ^** Division of Hemostasis and Thrombosis, Research Institute of Hematology, and ^§§ Department of Hematology, Jichi Medical School, Minamikawachi, Tochigi 329-04, Japan, the ^¶ Division of Cell Biology, Institute of Life Science, Soka University, 1-236 Tangi, Hachioji, Tokyo 192, Japan,  Katsuta Research Laboratory, Hitachi Koki Co., Ltd., Hitachinaka, Ibaraki 312, Japan, and the  Department of Internal Medicine, Jikei University School of Medicine, Tokyo 105, Japan

Sialyl-Le^x (sLe^x) antigen expression recognized by KM93 monoclonal antibody was significantly down-regulated during differentiation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in human pre-B lymphocytic leukemia cell line KM3. The sLe^x determinants were almost exclusively expressed on O-linked oligosaccharide chains of an O-glycosylated 150-kDa glycoprotein (gp150). A low shear force cell adhesion assay showed that TPA treatment significantly inhibited E-selectin-mediated cell adhesion. Transcript and/or enzyme activity levels of 13-fucosyltransferase, 23-sialyltransferase, 14-galactosyltransferase, and elongation 13-N-acetylglucosaminyltransferase did not correlate with sLe^x expression levels. However, transcript and enzyme activity levels of core 2 GlcNAc-transferase (C2GnT) were significantly down-regulated during TPA treatment. Following transfection and constitutive expression of full-length exogenous C2GnT transcript, C2GnT enzyme activities were maintained at high levels even after TPA treatment and down-regulation of cell surface sLe^x antigen expression by TPA was completely abolished. Furthermore, in the transfected cells, the KM93 reactivity of gp150 was not reduced by TPA treatment, and the inhibition of cell adhesion by TPA was also blocked. These results suggest that sLe^x expression is critically regulated by a single glycosyltransferase, C2GnT, during differentiation of KM3 cells.

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Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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