ZFIN ID: ZDB-PUB-090629-16
Zebrafish and Mouse α2,3-Sialyltransferases Responsible for Synthesizing GM4 Ganglioside
Chisada, S.I., Yoshimura, Y., Sakaguchi, K., Uemura, S., Go, S., Ikeda, K., Uchima, H., Matsunaga, N., Ogura, K., Tai, T., Okino, N., Taguchi, R., Inokuchi, J., and Ito, M.
Date: 2009
Source: The Journal of biological chemistry   284(44): 30534-30546 (Journal)
Registered Authors: Yoshimura, Yukihiro
Keywords: DEVELOPMENT DIFFERENTIATION, ENZYMES, ENZYMES/Lipid, GLYCOLIPIDS, GLYCOLIPIDS/Ganglioside, LIPID/Sphingolipids, MEMBRANE/Lipids
MeSH Terms:
  • Animals
  • Cloning, Molecular
  • DNA, Complementary
  • Embryo, Mammalian
  • Gangliosides/biosynthesis*
  • Mice
  • Mice, Knockout
  • Phylogeny*
  • RNA, Messenger/analysis
  • Sialyltransferases/genetics
  • Sialyltransferases/metabolism*
  • Tissue Distribution
  • Zebrafish
PubMed: 19542236 Full text @ J. Biol. Chem.
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ABSTRACT
We have previously reported that fish pathogens causing vibriosis specifically adhere to GM4 on the epithelial cells of fish intestinal tracts (Chisada et al. Biochem. Biophys. Res. Commun. 333, 367-373, 2005). To identify the gene encoding the enzyme for GM4 synthesis in the fish intestinal tract, a phylogenic tree of vertebrate ST3GalVs including Danio rerio and Oryzias latipes was generated, in which two putative sub-families of fish ST3GalVs were found. Two putative ST3GalVs of zebrafish (zST3GalV-1 and -2), each belonging to different sub-families, were cloned from the zebrafish cDNA library. Interestingly, zST3GalV-1 synthesized GM3 (NeuAcalpha2-3Galbeta1-4Glcbeta1-1'Cer) but not GM4 while zSTGalV-2 synthesized both gangliosides in vitro when expressed in CHO-K1 cells and RPMI1846 cells. Flow cytometric analysis using anti-GM4 antibody revealed that the transformation of RPMI1846 cells with zST3GalV-2 but not zST3GalV-1 cDNA increased the cell-surface expression of GM4. Whole mount in situ hybridization showed that the zST3GalV-2 transcript was strongly expressed in the gastrointestinal tract in 3 dpf embryos while zST3GalV-1 was expressed in the brain and esophagus but not gastrointestinal tract in 3 dpf embryos. On the other hand, we found that three isoforms of mouse ST3GalV (mST3GalV) having different N-terminal sequences can synthesize GM4 as well as GM3 in vitro when expressed in RPMI1846 and CHO-K1 cells. Furthermore, mST3GalV-knockout mice were found to lack GM4 synthase activity of the brain, and GM4 of the brain and erythrocytes, in contrast to wild-type mice. These results clearly indicate that zST3GalV-2 and mST3GalV are enzymes responsible for the synthesis of GM4 in zebrafish and mice, respectively.
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