Gene inactivation confirms the identity of enzymes involved in nematode phosphorylcholine-N-glycan synthesis

Katrina M. Houston, Ratneswary Sutharsan, C.N. Steiger, Harry Schachter, W. Harnett

Research output: Contribution to journalArticle

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Abstract

An unusual feature of nematodes is the covalent attachment of immunomodulatory phosphorylcholine (PC) moieties to N-type glycans. Our previous work on the filarial nematode glycoprotein ES-62 has enabled us to predict the identity of enzymes necessary for PC-N-glycan biosynthesis. Here, we addressed these predictions using gene knockout technology applied to C. elegans and present two pieces of confirmatory data. Employing a triple null mutant worm lacking all three genes that encode active UDP-N-acetyl-d-glucosamine: α-3-d-mannoside β1, 2-N-acetylglucosaminyltransferase I (GnT I) we have confirmed our earlier prediction that a crucial step in the generation of the substrate for PC transfer is addition of terminal GlcNAc to the α1-3-linked mannose residue of the glycan by GnT I. Second, by silencing genes responsible for expressing enzymes of the Kennedy pathway of phosphatidylcholine biosynthesis by RNA interference (RNAi), we have confirmed our belief for a role for diacylglycerol: choline phosphotransferase (CPT) in PC-N-glycan biosynthesis.
LanguageEnglish
Pages88-91
Number of pages4
JournalMolecular and Biochemical Parasitology
Volume157
Issue number1
Early online date9 Aug 2007
DOIs
Publication statusPublished - Jan 2008

Fingerprint

Phosphorylcholine
Gene Silencing
Polysaccharides
Enzymes
Mannosides
Gene Knockout Techniques
Uridine Diphosphate
Glucosamine
Diglycerides
Mannose
RNA Interference
Choline
Phosphatidylcholines
Glycoproteins
Phosphotransferases
Technology
Genes

Keywords

  • C. elegans
  • gene inactivation
  • N-glycan
  • phosphorylcholine
  • pharmacology

Cite this

Houston, Katrina M. ; Sutharsan, Ratneswary ; Steiger, C.N. ; Schachter, Harry ; Harnett, W. / Gene inactivation confirms the identity of enzymes involved in nematode phosphorylcholine-N-glycan synthesis. In: Molecular and Biochemical Parasitology. 2008 ; Vol. 157, No. 1. pp. 88-91.
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Gene inactivation confirms the identity of enzymes involved in nematode phosphorylcholine-N-glycan synthesis. / Houston, Katrina M.; Sutharsan, Ratneswary; Steiger, C.N.; Schachter, Harry; Harnett, W.

In: Molecular and Biochemical Parasitology, Vol. 157, No. 1, 01.2008, p. 88-91.

Research output: Contribution to journalArticle

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