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Effects of dibutyryl cAMP and bromodeoxyuridine on expression ofN-acetylglucosaminyltransferases III and V in GOTO neuroblastoma cells

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Abstract

The sugar chain structures of the cell surface change dramatically during cellular differentiation. A human neuroblastoma cell line, GOTO, is known to differentiate into neuronal cells and Schwannian cell-like cells on treatments with dibutyryl cAMP and bromodeoxyuridine, respectively. We have examined the expression of UDP-N-acetylglucosamine: β-d-mannoside β-1,4N-acetylglucosaminyltransferase III (GnT-III: EC 2.4.1.144) and UDP-N-acetylglucosamine: α-6-d-mannoside β-1,6N-acetylglucosaminyltransferase V (GnT-V: EC 2.4.1.155), two major branch forming enzymes inN-glycan synthesis, in GOTO cells on two distinct directions of differentiation.

In neuronal cell differentiation, GnT-III activity showed a slight increase during initial treatment with Bt2cAMP for 4 days and decreased drastically after the fourth day, but the mRNA level of GnT-III did not show a decrease but in fact a slight increase. GnT-V activity increased to approximately two- to three-fold the initial level with increasing mRNA level after 8 days, and lectin blot analysis showed an increase in reactivity toDatsura stramonium (DSA) of the immunoprecipitated neural cell adhesion molecule (NCAM). In Schwannian cell differentiation, the activity and mRNA level of GnT-III showed no significant change on treatment with BrdU. GnT-V activity also showed no change in spite of the gradual increase in the mRNA level. These results suggest that the activation of GnT-V during neuronal cell differentiation of GOTO cells might be a specific change for branch formation in N-glycans, and this affects the sugar chain structures of some glycoproteins such as NCAM.

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Abbreviations

GnT:

N-acetylglucosaminyltransferase

Bt2cAMP:

N 6,O 6-dibutyryl cAMP

BrdU:

bromodeoxyuridine

DSA:

Datsura stramonium

NCAM:

neural cell adhesion molecule

PAGE:

polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Department of Biochemistry, Osaka University Medical School, 2-2 Yamadaoka, 565, Suita, Osaka, Japan

    Yoshito Ihara, Atsushi Nishikawa & Naoyuki Taniguchi

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  1. Yoshito Ihara
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  2. Atsushi Nishikawa
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  3. Naoyuki Taniguchi
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Ihara, Y., Nishikawa, A. & Taniguchi, N. Effects of dibutyryl cAMP and bromodeoxyuridine on expression ofN-acetylglucosaminyltransferases III and V in GOTO neuroblastoma cells. Glycoconjugate J 12, 787–794 (1995). https://doi.org/10.1007/BF00731240

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