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Vol. 14, Issue 3, 1182-1194, March 2003
and
*Wellcome Centre for Molecular Parasitology, University of
Glasgow, The Anderson College, Glasgow G11 6NU, United Kingdom;
The survival of Trypanosoma brucei, the causative
agent of Sleeping Sickness and Nagana, is facilitated by the expression of a dense surface coat of glycosylphosphatidylinositol
(GPI)-anchored proteins in both its mammalian and tsetse fly hosts. We
have characterized T. brucei GPI8, the gene encoding the
catalytic subunit of the GPI:protein transamidase complex that adds
preformed GPI anchors onto nascent polypeptides. Deletion of
GPI8 (to give Wellcome Trust Laboratories for Molecular
Parasitology, Department of Biological Sciences and Centre for
Molecular Microbiology and Infection, Imperial College, London SW7 2AY,
United Kingdom; and Division of Infection and
Immunity, Institute of Biomedical and Life Sciences, University of
Glasgow, Glasgow G12 8QQ, United Kingdom
gpi8) resulted in the
absence of GPI-anchored proteins from the cell surface of procyclic
form trypanosomes and accumulation of a pool of non-protein-linked GPI
molecules, some of which are surface located. Procyclic
gpi8, while viable in culture, were unable to
establish infections in the tsetse midgut, confirming that GPI-anchored
proteins are essential for insect-parasite interactions. Applying
specific inducible GPI8 RNAi with bloodstream form
parasites resulted in accumulation of unanchored variant surface
glycoprotein and cell death with a defined multinuclear,
multikinetoplast, and multiflagellar phenotype indicative of a block in
cytokinesis. These data show that GPI-anchored proteins are essential
for the viability of bloodstream form trypanosomes even in the absence
of immune challenge and imply that GPI8 is important for proper cell
cycle progression.
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