Molecular characterization of the cis-prenyltransferase of Giardia lamblia.

Journal Article (Journal Article)

Giardia lamblia, the protist that causes diarrhea, makes an Asn-linked-glycan (N-glycan) precursor that contains just two sugars (GlcNAc(2)) attached by a pyrophosphate linkage to a polyprenol lipid. Because the candidate cis-prenyltransferase of Giardia appears to be more similar to bacterial enzymes than to those of most eukaryotes and because Giardia is missing a candidate dolichol kinase (ortholog to Saccharomyces cerevisiae SEC59 gene product), we wondered how Giardia synthesizes dolichol phosphate (Dol-P), which is used to make N-glycans and glycosylphosphatidylinositol (GPI) anchors. Here we show that cultured Giardia makes an unsaturated polyprenyl pyrophosphate (dehydrodolichol), which contains 11 and 12 isoprene units and is reduced to dolichol. The Giardia cis-prenyltransferase that we have named Gl-UPPS because the enzyme primarily synthesizes undecaprenol pyrophosphate is phylogenetically related to those of bacteria and Trypanosoma rather than to those of other protists, metazoans and fungi. In transformed Saccharomyces, the Giardia cis-prenyltransferase also makes a polyprenol containing 11 and 12 isoprene units and supports normal growth, N-glycosylation and GPI anchor synthesis of a rer2Delta, srt1Delta double-deletion mutant. Finally, despite the absence of an ortholog to SEC59, Giardia has cytidine triphosphate-dependent dolichol kinase activity. These results suggest that the synthetic pathway for Dol-P is conserved in Giardia, even if some of the important enzymes are different from those of higher eukaryotes or remain unidentified.

Full Text

Duke Authors

Cited Authors

  • GrabiƄska, KA; Cui, J; Chatterjee, A; Guan, Z; Raetz, CRH; Robbins, PW; Samuelson, J

Published Date

  • July 2010

Published In

Volume / Issue

  • 20 / 7

Start / End Page

  • 824 - 832

PubMed ID

  • 20308470

Pubmed Central ID

  • PMC2900897

Electronic International Standard Serial Number (EISSN)

  • 1460-2423

Digital Object Identifier (DOI)

  • 10.1093/glycob/cwq036


  • eng

Conference Location

  • England