ZFIN ID: ZDB-PUB-120223-32
Identification and biochemical characterization of two functional CMP-sialic acid synthetases in Danio rerio
Schaper, W., Bentrop, J., Ustinova, J., Blume, L., Kats, E., Tiralongo, J., Weinhold, B., Bastmeyer, M., and Muenster-Kuehnel, A.
Date: 2012
Source: The Journal of biological chemistry   287(16): 13239-13248 (Journal)
Registered Authors: Bastmeyer, Martin, Bentrop, Joachim
Keywords: enzymes, glycobiology, glycosylation, sialic acid, zebrafish, CMP-sialic acid synthetase
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Cell Line, Tumor
  • Cell Nucleus/enzymology
  • Evolution, Molecular*
  • Gene Expression Regulation, Developmental/physiology
  • Gene Expression Regulation, Enzymologic/physiology
  • Glycosylation
  • Mice
  • Molecular Sequence Data
  • N-Acetylneuraminic Acid/metabolism
  • N-Acylneuraminate Cytidylyltransferase/chemistry
  • N-Acylneuraminate Cytidylyltransferase/genetics*
  • N-Acylneuraminate Cytidylyltransferase/metabolism
  • NIH 3T3 Cells
  • RNA, Messenger/genetics
  • Substrate Specificity/physiology
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed: 22351762 Full text @ J. Biol. Chem.

Sialic acids (Sia) form the non-reducing end of the bulk of cell surface expressed glycoconjugates. They are, therefore, major elements in intercellular communication processes. Addition of Sia to glycoconjugates requires metabolic activation to CMP-Sia, catalyzed by CMP-Sia synthetase (CMAS). This highly conserved enzyme is located in the cell nucleus in all vertebrates investigated to date, but its nuclear function remains elusive. Here, we describe the identification and characterization of two Cmas enzymes in Danio rerio (dreCmas), one of which exclusively localized in the cytosol. We show that the two cmas genes most likely originated from the third whole genome duplication which occurred at the base of teleost radiation. cmas paralogues were maintained in fishes of the Otocephala clade, while one copy got subsequently lost in Euteleostei (e.g. rainbow trout). In zebrafish, the two genes exhibited a distinct spatial expression pattern. The products of these genes (dreCmas1 and dreCmas2) diverged not only with respect to subcellular localization but also in substrate specificity. Nuclear dreCmas1 favored N-acetylneuraminic acid (Neu5Ac), while the cytosolic dreCmas2 showed highest affinity for 5-deamino-neuraminic acid (KDN). The subcellular localization was confirmed for the endogenous enzymes in fractionated zebrafish lysates. Nuclear entry of dreCmas1 was mediated by a bipartite nuclear localization signal, which seemed irrelevant for other enzymatic functions. With the current demonstration that in zebrafish two subfunctionalized cmas paralogues co-exist, we introduce a novel and unique model to detail the roles that CMAS has in the nucleus and in the sialylation pathways of animal cells.