Mammalian cytidine 5'-monophosphate N-acetylneuraminic acid synthetase: A nuclear protein with evolutionarily conserved structural motifs

A. K. Munster, M. Eckhardt, B. Potvin, M. Muhlenhoff, Pamela Stanley, R. Gerardy- Schahn

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

Sialic acids of cell surface glycoproteins and glycolipids play a pivotal role in the structure and function of animal tissues. The pattern of cell surface sialylation is species and tissue-specific, is highly regulated during embryonic development, and changes with stages of differentiation. A prerequisite for the synthesis of sialylated glycoconjugates is the activated sugar-nucleotide cytidine 5'-monophosphate N-acetylneuraminic acid (CMP- Neu5Ac), which provides a substrate for Golgi sialyltransferases. Although a mammalian enzymatic activity responsible for the synthesis of CMP-Neu5Ac has been described and the enzyme has been purified to near homogeneity, sequence information is restricted to bacterial CMP-Neu5Ac synthetases. In this paper, we describe the molecular characterization, functional expression, and subcellular localization of murine CMP-Neu5Ac synthetase. Cloning was achieved by complementation of the Chinese hamster ovary lec32 mutation that causes a deficiency in CMP-Neu5Ac synthetase activity. A murine cDNA encoding a protein of 432 amino acids rescued the lec32 mutation and also caused polysialic acid to be expressed in the capsule of the CMP-Neu5Ac synthetase negative Escherichia coli mutant EV5. Three potential nuclear localization signals were found in the murine synthetase, and immunofluorescence studies confirmed predominantly nuclear localization of an N-terminally Flag-tagged molecule. Four stretches of amino acids that occur in the N-terminal region are highly conserved in bacterial CMP-Neu5Ac synthetases, providing evidence for an ancestral relationship between the sialylation pathways of bacterial and animal cells.

Original languageEnglish (US)
Pages (from-to)9140-9145
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number16
DOIs
StatePublished - Aug 4 1998

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Cytidine Monophosphate N-Acetylneuraminic Acid
Ligases
Nuclear Proteins
Sialyltransferases
Sialic Acids
Animal Structures
Amino Acids
Nuclear Localization Signals
Mutation
Glycoconjugates
Glycolipids
Membrane Glycoproteins
Cricetulus
cytidine-5'-monophosphosialic acid
Capsules
Embryonic Development
Fluorescent Antibody Technique
Organism Cloning
Ovary
Nucleotides

Keywords

  • Nuclear localization
  • Nucleotidyltransferases
  • Polysialic acid
  • Sialic acid

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Mammalian cytidine 5'-monophosphate N-acetylneuraminic acid synthetase : A nuclear protein with evolutionarily conserved structural motifs. / Munster, A. K.; Eckhardt, M.; Potvin, B.; Muhlenhoff, M.; Stanley, Pamela; Gerardy- Schahn, R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, No. 16, 04.08.1998, p. 9140-9145.

Research output: Contribution to journalArticle

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