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Glycobiology Advance Access originally published online on October 13, 2004
Glycobiology 2005 15(3):271-280; doi:10.1093/glycob/cwi005
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Glycobiology vol. 15 no. 3 © Oxford University Press 2005; all rights reserved.

Not core 2 ß1,6-N-acetylglucosaminyltransferase-2 or -3 but -1 regulates sialyl-Lewis x expression in human precursor B cells

Jiro Kikuchi2, Hirotaka Shinohara2,3, Chizu Nonomura2,4, Hidenobu Ando2, Shizuka Takaku2, Hisao Nojiri3 and Mitsuru Nakamura1,2,4

2 Cell Regulation Analysis Team, Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Central-4, 1-1-1 Higashi, Tsukuba, 305-8562, Japan; 3 Department of Biochemical Oncology, School of Pharmaceutical Sciences, Teikyo University, Sagamiko 199-0195, Japan; and 4 Core Research for Evolution Science and Technology (CREST) of Japan Science and Technology Corporation, Kawaguchi, 332-0012, Japan

1 To whom correspondence should be addressed; e-mail: owlm.nakamura{at}aist.go.jp

Received on September 3, 2004; revised on October 7, 2004; accepted on October 8, 2004

Sialyl-Lewis x (sLeX), one of the major selectin ligands, is expressed on T and B cells in a differentiation or activation stage-specific manner. We have demonstrated before that sLeX expression and core 2 ß1,6-N-acetylglucosaminyltransferase (C2GnT) were simultaneously regulated during precursor B (pre-B) cell differentiation. Three C2GnT family genes, designated C2GnT-1, -2, and -3, were previously identified, but their roles have not been fully examined. In this study, we have investigated the roles of C2GnTs in the regulation of sLeX expression level during pre-B cell differentiation comparing with {alpha}1,3fucosyltransferase-VII (FucT-VII) and {alpha}2,3sialyltransferase-IV (ST3Gal-IV). Overexpression of not FucT-VII and ST3Gal-IV but C2GnT-1 blocked the down-regulation of sLeX expression by differentiation induction. Neither C2GnT-2 nor -3 but C2GnT-1 transcript was mainly expressed in B lineage cell lines and bone marrow–derived B lineage cells. Significant down-regulation of C2GnT-1 of the three C2GnTs was observed in KM3 cells during differentiation. The expression of C2GnT-1 correlated well to sLeX expression and differentiation stage. Furthermore, introduction of short interfering RNA against C2GnT-1 markedly reduced C2GnT-1 expression and resulted in down-regulation of sLeX expression. These results suggest that not the other glycosyltransferases but C2GnT-1 regulates sLeX expression level during differentiation of pre-B cells, providing the cells with substrate of sLeX structure biosynthesis.

Key words: siayl-Lewis x / core 2 GlcNAc transferase / precursor-B cell / differentiation / short intefering RNA


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