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Molecular Characterization of Toxoplasma gondii Formin 3, an Actin Nucleator Dispensable for Tachyzoite Growth and Motility

Wassim Daher, Natacha Klages, Marie-France Carlier, Dominique Soldati-Favre
Wassim Daher
aDepartment of Microbiology and Molecular Medicine, CMU, University of Geneva, Geneva, Switzerland
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Natacha Klages
aDepartment of Microbiology and Molecular Medicine, CMU, University of Geneva, Geneva, Switzerland
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Marie-France Carlier
bDynamique du Cytosquelette, Laboratoire d'Enzymologie et Biochimie Structurales UPR A 9063, CNRS, Gif sur Yvette, France
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Dominique Soldati-Favre
aDepartment of Microbiology and Molecular Medicine, CMU, University of Geneva, Geneva, Switzerland
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DOI: 10.1128/EC.05192-11
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ABSTRACT

Toxoplasma gondii belongs to the phylum Apicomplexa, a group of obligate intracellular parasites that rely on gliding motility to enter host cells. Drugs interfering with the actin cytoskeleton block parasite motility, host cell invasion, and egress from infected cells. Myosin A, profilin, formin 1, formin 2, and actin-depolymerizing factor have all been implicated in parasite motility, yet little is known regarding the importance of actin polymerization and other myosins for the remaining steps of the parasite lytic cycle. Here we establish that T. gondii formin 3 (TgFRM3), a newly described formin homology 2 domain (FH2)-containing protein, binds to Toxoplasma actin and nucleates rabbit actin assembly in vitro. TgFRM3 expressed as a transgene exhibits a patchy localization at several distinct structures within the parasite. Disruption of the TgFRM3 gene by double homologous recombination in a ku80-ko strain reveals no vital function for tachyzoite propagation in vitro, which is consistent with its weak level of expression in this life stage. Conditional stabilization of truncated forms of TgFRM3 suggests that different regions of the molecule contribute to distinct localizations. Moreover, expression of TgFRM3 lacking the C-terminal domain severely affects parasite growth and replication. This work provides a first insight into how this specialized formin, restricted to the group of coccidia, completes its actin-nucleating activity.

FOOTNOTES

    • Received 29 July 2011.
    • Accepted 19 December 2011.
    • Accepted manuscript posted online 30 December 2011.
  • Supplemental material for this article may be found at http://dx.doi.org/10.1128/EC.05192-11.

  • Copyright © 2012, American Society for Microbiology. All Rights Reserved.
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Molecular Characterization of Toxoplasma gondii Formin 3, an Actin Nucleator Dispensable for Tachyzoite Growth and Motility
Wassim Daher, Natacha Klages, Marie-France Carlier, Dominique Soldati-Favre
Eukaryotic Cell Feb 2012, 11 (3) 343-352; DOI: 10.1128/EC.05192-11

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Molecular Characterization of Toxoplasma gondii Formin 3, an Actin Nucleator Dispensable for Tachyzoite Growth and Motility
Wassim Daher, Natacha Klages, Marie-France Carlier, Dominique Soldati-Favre
Eukaryotic Cell Feb 2012, 11 (3) 343-352; DOI: 10.1128/EC.05192-11
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