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Structure–function analysis of the interaction between Bax and the cytomegalovirus-encoded protein vMIA

Oncogene volume 26, pages 7067–7080 (2007)Cite this article

Abstract

The viral mitochondrial inhibitor of apoptosis (vMIA) encoded by the human cytomegalovirus exerts cytopathic effects and neutralizes the proapoptotic endogenous Bcl-2 family member Bax by recruiting it to mitochondria, inducing its oligomerization and membrane insertion. Using a combination of computational modeling and mutational analyses, we addressed the structure–function relationship of the molecular interaction between the protein Bax and the viral antiapoptotic protein vMIA. We propose a model in which vMIA exhibits an overall fold similar to Bcl-XL. In contrast to Bcl-XL, however, this predicted conformation of vMIA does not bind to the BH3 domain of Bax and rather engages in electrostatic interactions that involve a stretch of amino acids between the BH3 and BH2 domains of Bax and an α-helical domain located within the previously defined Bax-binding domain of vMIA, between the putative BH1-like and BH2-like domains. According to this model, vMIA is likely to bind Bax preferentially in its membrane-inserted conformation. The capacity of vMIA to cause fragmentation of the mitochondrial network and disorganization of the actin cytoskeleton is independent of its Bax-binding function. We found that Δ131–147 vMIA mutant, which lacks both the Bax-binding function and cell-death suppression but has intact mitochondria-targeting capacity, is similar to vMIA in its ability to disrupt the mitochondrial network and to disorganize the actin cytoskeleton. vMIAΔ131–147 is a dominant-negative inhibitor of the antiapoptotic function of wild-type vMIA. Our experiments with vMIAΔ131–147 suggest that vMIA forms homo-oligomers, which may engage in cooperative and/or multivalent interactions with Bax, leading to its functional neutralization.

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Abbreviations

BBD:

Bax-binding domain

CMV:

human cytomegalovirus

CHAPS:

3-((cholamidopropyl)dimethylammonio)-1-propane sulfonate

DiOC6(3):

3,3′-dihexyloxacarbocyanine iodide

GFP:

green fluorescent protein

MLS:

mitochondrial localization sequence

MOMP:

mitochondrial outer membrane permeabilization

PI:

propidium iodide

STS:

staurosporine

TMRM:

tetra methyl rhodamine methyl ester

vMIA:

viral mitochondria-localized inhibitor of apoptosis

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Acknowledgements

We thank Dr Matsuyama (Medical College of Wisconsin, Milwaukee, WI, USA) for Bax expression plasmid. GK is supported by Ligue Nationale contre le cancer, European Community (Active p53, RIGHT), Agence National pour la Recherche contre le Sida (ANRS) and Sidaction.

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Authors and Affiliations

  1. INSERM, U848, Pavillion de Recherche 1, Villejuif, France

    A-L Pauleau, N Larochette, S R Scholz, D Poncet, N Zamzami & G Kroemer

  2. Institut Gustave Roussy, Villejuif, France

    A-L Pauleau, N Larochette, S R Scholz, D Poncet, N Zamzami & G Kroemer

  3. Faculté de Médecine – Université Paris-Sud XI, Villejuif, France

    A-L Pauleau, N Larochette, S R Scholz, D Poncet, N Zamzami & G Kroemer

  4. Lead generation, AstraZeneca R&D Mölndal, Pepparedsleden 1, Mölndal, Sweden

    F Giordanetto

  5. ImmunoGen Inc., Cambridge, MA, USA

    V S Goldmacher

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  1. A-L Pauleau
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Correspondence to G Kroemer.

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Pauleau, AL., Larochette, N., Giordanetto, F. et al. Structure–function analysis of the interaction between Bax and the cytomegalovirus-encoded protein vMIA. Oncogene 26, 7067–7080 (2007). https://doi.org/10.1038/sj.onc.1210511

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