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Title

Trans-genera reconstitution and complementation of an adhesion complex in Toxoplasma gondii

Authors
Huynh, My-Hang
Opitz, Corinna
Kwok, Lai-Yu
Tomley, Fiona M
Carruthers, Vern B
Published in Cellular Microbiology. 2004, vol. 6, no. 8, p. 771-782
Abstract Eimeria tenella and Toxoplasma gondii are obligate intracellular parasites belonging to the phylum Apicomplexa. In T. gondii, the microneme protein TgMIC2 contains two well-defined adhesive motifs and is thought to be a key participant in the attachment and invasion of host cells. However, several attempts by different laboratories to generate a knockout (KO) of TgMIC2 have failed, implying that TgMIC2 is an essential gene. As Eimeria and Toxoplasma utilize the same mechanisms of invasion and have highly conserved adhesive proteins, we hypothesized that the orthologous molecule in Eimeria, EtMIC1, could functionally substitute in Toxoplasma to allow a knockout of TgMIC2. TgMIC2 is partnered with a protein called TgM2AP, which corresponds to EtMIC2 in Eimeria. Because the activity of TgMIC2 is most likely tightly linked to its association with TgM2AP, it was thought that the activity of EtMIC1 might similarly require its partner EtMIC2. EtMIC1 and EtMIC2 were introduced into T. gondii, and the presence of EtMIC1 allowed the first knockout clone of TgMIC2 to be obtained. The TgMIC2 KO resulted in significantly decreased numbers of invaded parasites compared to the parental clone. In the absence of TgMIC2, TgM2AP was incorrectly processed and mistargeted to the parasitophorous vacuole instead of the micronemes. These findings indicate that the EtMIC1 can compensate for the essential requirement of TgMIC2, but it cannot fully functionally substitute for TgMIC2 in the invasion process or for supporting the correct maturation and targeting of TgM2AP.
Keywords Amino Acid SequenceAnimalsCell Adhesion/geneticsCell Adhesion Molecules/genetics/metabolismClone CellsDisease Models, AnimalEimeria/geneticsGenes, ProtozoanGenetic Complementation TestHumansMembrane Proteins/genetics/metabolismMiceMice, Inbred BALB CMolecular Sequence DataProtozoan Proteins/genetics/metabolismSequence AlignmentToxoplasma/genetics/pathogenicityTransfectionVirulence/genetics
Identifiers
PMID: 15236644
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Article (Author postprint) (717 Kb) - public document Free access
Structures
Research group Biologie d'un parasite intracellulaire obligatoire (773)
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HUYNH, My-Hang et al. Trans-genera reconstitution and complementation of an adhesion complex in Toxoplasma gondii. In: Cellular Microbiology, 2004, vol. 6, n° 8, p. 771-782. https://archive-ouverte.unige.ch/unige:78034

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Deposited on : 2015-12-02

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