title: Dual acylation accounts for the localization of {alpha}19-giardin in the ventral flagellum pair of Giardia lamblia.
abstract: A Giardia-specific protein family denominated as alpha-giardins, represents the major protein component, besides tubulin, of the cytoskeleton of the human pathogenic parasite Giardia lamblia. One of its members, alpha19-giardin, carries an N-terminal sequence extension of MGCXXS, which in many proteins serves as a target for dual lipid conjugation: myristoylation at the glycine residue after removal of the methionine and palmitoylation at the cysteine residue. As the first experimental evidence of a lipid modification, we found alpha19-giardin to be associated with the membrane fraction of disrupted trophozoites. After heterologous coexpression of alpha19-giardin with giardial N-myristoyltransferase (NMT) in Escherichia coli, we found the protein in a myristoylated form. Additionally, after heterologous expression together with the palmitoyl transferase Pfa3 in Saccharomyces cerevisiae, alpha19-giardin associates with the membrane of the main vacuole. Immunocytochemical colocalization studies on wild-type Giardia trophozoites with tubulin provide evidence that alpha19-giardin exclusively localizes to the ventral pair of the giardial flagella. A mutant in which the putatively myristoylated N-terminal glycine residue was replaced by alanine lost this specific localization. Our findings suggest that the dual lipidation of alpha19-giardin is responsible for its specific flagellar localization.
authors.affiliation: Faculty of Biology/Chemistry, Barbarastrasse 13, D-49069 Osnabrueck, Germany.
authors.forename: Mirela
authors.initials: M
authors.lastname: Saric
authors.forename: Anke
authors.initials: A
authors.lastname: Vahrmann
authors.forename: Daniela
authors.initials: D
authors.lastname: Niebur
authors.forename: Verena
authors.initials: V
authors.lastname: Kluempers
authors.forename: Adrian B
authors.initials: AB
authors.lastname: Hehl
authors.forename: Henning
authors.initials: H
authors.lastname: Scholze
mesh_terms: D000215:Acylation; D003598:Cytoskeletal Proteins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D005407:Flagella / Q000378:metabolism*; D016829:Giardia lamblia / Q000378:metabolism*; D018613:Microscopy, Confocal; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D015800:Protozoan Proteins; C038207:giardin protein, Giardia lamblia
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