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Rapid control of protein level in the apicomplexan Toxoplasma gondii

Nature Methods volume 4, pages 1003–1005 (2007)Cite this article

An Erratum to this article was published on 01 January 2008

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Abstract

Analysis of gene function in apicomplexan parasites is limited by the absence of reverse genetic tools that allow easy and rapid modulation of protein levels. The fusion of a ligand-controlled destabilization domain (ddFKBP) to a protein of interest enables rapid and reversible protein stabilization in T. gondii. This allows an efficient functional analysis of proteins that have a dual role during host cell invasion and/or intracellular growth of the parasite.

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Figure 1: Rapid regulation of proteins fused to ddFKBP.
Figure 2: Functional analysis of essential proteins using the ddFKBP system.

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Change history

  • 29 November 2007

    Note: In the version of this article initially published, the labels in the bottom right image of Figure 2b were incorrect. The correct labels are shown in the image below. The error has been corrected in the HTML and PDF versions of the article.

Notes

  1. Note: In the version of this article initially published, the labels in the bottom right image of Figure 2b were incorrect. The correct labels are shown in the image below. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank B. Striepen (University of Georgia), D.L. Sibley (Washington University), V.B. Carruthers (Johns Hopkins School of Public Health), C.J. Beckers (The University of North Carolina) and D. Soldati (University of Geneva) for sharing reagents. We thank K. Matuschewski, G. Langsley and D. Goldberg for critically reading the manuscript, M. Rauch for technical assistance, and M. Kudryashev for help with image analysis. This work was funded by the BioFuture-Programm (grant 0311897) of the German ministry of science and education (BMBF).

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Author notes

  1. Angelika Herm-Götz, Carolina Agop-Nersesian and Sylvia Münter: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Parasitology, Hygieneinstitut, University Hospital Heidelberg, Im Neuenheimer Feld 324, Heidelberg, D-69120, Germany

    Angelika Herm-Götz, Carolina Agop-Nersesian, Sylvia Münter, Friedrich Frischknecht & Markus Meissner

  2. Department of Chemistry and Systems Biology, Stanford University, 269 Campus Drive, Stanford, 94305, California, USA

    Joshua S Grimley & Thomas J Wandless

Authors
  1. Angelika Herm-Götz
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  2. Carolina Agop-Nersesian
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  3. Sylvia Münter
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  4. Joshua S Grimley
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  5. Thomas J Wandless
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  7. Markus Meissner
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Contributions

A.H.-G generated and analyzed parasites expressing ddFKBP-YFP,GFP-ddFKBP and ddFKBP-MyoA. C.A.-N. generated and analyzed parasites expressing ddFKBP-Rab11ADN, ddFKBP-YPT1 and ddFKBP-YPT1DN. S.M. performed live cell imaging. J.S.G. synthesized Shld1. T.J.W. was involved in designing and discussing this study. F.F. and M.M. initiated and guided this study.

Corresponding author

Correspondence to Markus Meissner.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Methods (PDF 4132 kb)

Supplementary Movie 1

Upregulation of DD-YFP upon addition of Shld1 (MOV 3185 kb)

Supplementary Movie 2

Downregulation of DD-YFP upon removal of Shld1 (MOV 2698 kb)

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Herm-Götz, A., Agop-Nersesian, C., Münter, S. et al. Rapid control of protein level in the apicomplexan Toxoplasma gondii. Nat Methods 4, 1003–1005 (2007). https://doi.org/10.1038/nmeth1134

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