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Conditional and reversible disruption of essential herpesvirus proteins

Nature Methods volume 6, pages 577–579 (2009)Cite this article

Abstract

Elucidating the function of essential proteins of complex pathogenic viruses is impeded by a paucity of complementing systems. By fusing a destabilizing domain of the FK506-binding protein to essential cytomegalovirus proteins, we generated virus mutants in which amounts of fusion proteins and viral growth can be regulated by the synthetic ligand shield-1. This conditional approach will greatly facilitate the analysis of gene functions of herpesviruses and viruses of other families.

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Figure 1: Ligand-dependent expression of CMV ddFKBP fusion proteins after infection, and tunability of plaque formation.
Figure 2: Growth kinetics of the CMV mutants in the presence or absence of shield-1.

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Acknowledgements

We thank T. Wandless (Stanford University) for providing the shield-1 ligand and plasmids encoding ddFKBP and S. Jonjic (University of Rijeka) for monoclonal antibody Croma 101. We thank P. Kay-Jackson for critical reading of the manuscript. This work was in part supported by a grant of the Deutsche Forschungsgemeinschaft (collaborative research grant 587, individual project A13).

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Authors and Affiliations

  1. Department of Virology, Hannover Medical School, Hannover, Germany

    Mandy Glaß, Andreas Busche, Karen Wagner, Martin Messerle & Eva Maria Borst

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  1. Mandy Glaß
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  2. Andreas Busche
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  4. Martin Messerle
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Contributions

E.M.B. conceived the study and together with M.M. designed experiments and wrote the manuscript. E.M.B., K.W., M.G. and A.B. performed experiments and analyzed data. M.G. assisted in writing the manuscript.

Corresponding author

Correspondence to Martin Messerle.

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Supplementary Figures 1–5, Supplementary Table 1, Supplementary Note 1 (PDF 5693 kb)

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Glaß, M., Busche, A., Wagner, K. et al. Conditional and reversible disruption of essential herpesvirus proteins. Nat Methods 6, 577–579 (2009). https://doi.org/10.1038/nmeth.1346

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