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The orf virus inhibitor of apoptosis functions in a Bcl-2-like manner, binding and neutralizing a set of BH3-only proteins and active Bax

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Abstract

We have previously shown that the Orf virus protein, ORFV125, is a potent inhibitor of the mitochondrial pathway of apoptosis and displays rudimentary sequence similarities to cellular anti-apoptotic Bcl-2 proteins. Here we investigate the proposal that ORFV125 acts in a Bcl-2-like manner to inhibit apoptosis. We show that the viral protein interacted with a range of BH3-only proteins (Bik, Puma, DP5, Noxa and all 3 isoforms of Bim) and neutralized their pro-apoptotic activity. In addition, ORFV125 bound to the active, but not the inactive, form of Bax, and reduced the formation of Bax dimers. Mutation of specific amino acids in ORFV125 that are conserved and functionally important in mammalian Bcl-2 family proteins led to loss of both binding and inhibitory functions. We conclude that ORFV125’s mechanism of action is Bcl-2-like and propose that the viral protein’s combined ability to bind to a range of BH3-only proteins as well as the active form of Bax provides significant protection against apoptosis. Furthermore, we demonstrate that the binding profile of ORFV125 is distinct to that of other poxviral Bcl-2-like proteins.

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Abbreviations

Bcl-2:

B-cell leukemia/lymphoma 2

BH:

Bcl-2 homology

ORFV:

Orf virus

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Acknowledgments

This work was supported by the Health Research Council of New Zealand and the University of Otago. We wish to thank David Huang and Jamie Fletcher (The Walter and Eliza Hall Institute of Medical Research (WEHI), Australia) for providing information and a range of plasmids expressing Bcl-2 family members. We are grateful to Jerry Adams (WEHI, Australia) for critical reading of the manuscript, Catherine Day and Fabienne Lecomte (University of Otago, New Zealand) for helpful discussions and Ellena Whelan (University of Otago) for technical support.

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Authors and Affiliations

  1. Virus Research Unit, Department of Microbiology and Immunology, University of Otago, P.O. Box 56, Dunedin, New Zealand

    Dana Westphal, Merilyn H. Hibma, Norihito Ueda, Stephen B. Fleming & Andrew A. Mercer

  2. Department of Biochemistry, University of Otago, Dunedin, New Zealand

    Elizabeth C. Ledgerwood

  3. The National School of Pharmacy, University of Otago, Dunedin, New Zealand

    Joel D. A. Tyndall

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  1. Dana Westphal
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  2. Elizabeth C. Ledgerwood
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  6. Stephen B. Fleming
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Correspondence to Andrew A. Mercer.

Additional information

Elizabeth C. Ledgerwood and Andrew A. Mercer contributed equally to this work.

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Westphal, D., Ledgerwood, E.C., Tyndall, J.D.A. et al. The orf virus inhibitor of apoptosis functions in a Bcl-2-like manner, binding and neutralizing a set of BH3-only proteins and active Bax. Apoptosis 14, 1317–1330 (2009). https://doi.org/10.1007/s10495-009-0403-1

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