title: Stage-specific depletion of myosin A supports an essential role in motility of malarial ookinetes.
abstract: Functional analysis of Plasmodium genes by classical reverse genetics is currently limited to mutants that are viable during erythrocytic schizogony, the pathogenic phase of the malaria parasite where transfection is performed. Here, we describe a conceptually simple experimental approach to study the function of genes essential to the asexual blood stages in a subsequent life cycle stage by a promoter-swap approach. As a proof of concept we targeted the unconventional class XIV myosin MyoA, which is known to be required for Toxoplasma gondii tachyzoite locomotion and host cell invasion. By placing the corresponding Plasmodium berghei gene, PbMyoA, under the control of the apical membrane antigen 1 (AMA1) promoter, expression in blood stages is maintained but switched off during transmission to the insect vector, i.e. ookinetes. In those mutant ookinetes gliding motility is entirely abolished resulting in a complete block of life cycle progression in Anopheles mosquitoes. Similar approaches should permit the analysis of gene function in the mosquito forms that are shared with the erythrocytic stages of the malaria parasite.
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece.
authors: Inga
authors: I
authors: Siden-Kiamos
authors: Markus
authors: M
authors: Ganter
authors: Andreas
authors: A
authors: Kunze
authors: Marion
authors: M
authors: Hliscs
authors: Marion
authors: M
authors: Steinbüchel
authors: Jacqueline
authors: J
authors: Mendoza
authors: Robert E
authors: RE
authors: Sinden
authors: Christos
authors: C
authors: Louis
authors: Kai
authors: K
authors: Matuschewski
mesh_terms: D000818:Animals; D000852:Anopheles / Q000469:parasitology; D000953:Antigens, Protozoan / Q000235:genetics / Q000378:metabolism*; D005260:Female; D017353:Gene Deletion; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D005816:Genetic Complementation Test; D008124:Locomotion; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008845:Microinjections; D009218:Myosins / Q000235:genetics / Q000378:metabolism*; D033661:Oocysts / Q000378:metabolism; D010962:Plasmodium berghei / Q000235:genetics; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D034101:Sporozoites / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D014123:Toxoplasmosis / Q000469:parasitology; D014162:Transfection
chemical_list: D000953:Antigens, Protozoan; D008565:Membrane Proteins; C435214:MyoA protein, Plasmodium berghei; D015800:Protozoan Proteins; C090077:apical membrane antigen I, Plasmodium; D009218:Myosins