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A deneddylase encoded by Epstein–Barr virus promotes viral DNA replication by regulating the activity of cullin-RING ligases

Nature Cell Biology volume 12, pages 351–361 (2010)Cite this article

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Abstract

The large tegument proteins of herpesviruses encode conserved cysteine proteases of unknown function. Here we show that BPLF1, the Epstein–Barr-virus-encoded member of this protease family, is a deneddylase that regulates virus production by modulating the activity of cullin-RING ligases (CRLs). BPLF1 hydrolyses NEDD8 conjugates in vitro, acts as a deneddylase in vivo, binds to cullins and stabilizes CRL substrates. Expression of BPLF1 alone or in the context of the productive virus cycle induces accumulation of the licensing factor CDT1 and deregulates S-phase DNA synthesis. Inhibition of BPLF1 during the productive virus cycle prevents cellular DNA re-replication and inhibits virus replication. Viral DNA synthesis is restored by overexpression of CDT1. Homologues encoded by other herpesviruses share the deneddylase activity. Thus, these enzymes are likely to have a key function in the virus life cycle by inducing a replication-permissive S-phase-like cellular environment.

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Figure 1: BPLF1 is a NEDD8-specific deconjugase.
Figure 2: BPLF1 deneddylates cullins and induces the accumulation of CRL substrates.
Figure 3: Expression of a catalytically active BPLF1 induces accumulation of cells with a DNA content of at least 4N.
Figure 4: Cell cycle deregulation in cells expressing BPLF1.
Figure 5: Activation of the EBV productive cycle is associated with cellular DNA re-replication.
Figure 6: Expression of BPLF1 is required for efficient EBV DNA replication.
Figure 7: Overexpression of CDT1 promotes EBV DNA replication.
Figure 8: Deneddylase activity is shared by BPLF1 homologues encoded by other herpesviruses.

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Acknowledgements

We thank Ron T. Hay, Frauke Melchior, Zhen-Quian-Pan, Dong-Er Zhang, Chiba Chikatumi and Jürgen Haas for providing plasmids, antibodies and technical advice. This study was supported by grants awarded by the Swedish Cancer Society, the Swedish Medical Research Council, the Karolinska Institutet, Stockholm, Sweden, and by the European Community Integrated Project INCA, contract no. LSHC-CT-2005-018704, and Network of Excellence RUBICON, contract no. LSG-CT-2005-018683. S.H. is supported by a fellowship from the European Community Marie Curie Early Training Network UbiRegulators contract no. MRTN-CT-2006-034555.

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  1. Department of Cell and Molecular Biology, Karolinska Institutet, Box 285, S-17177, Stockholm, Sweden

    Stefano Gastaldello, Sebastian Hildebrand, Omid Faridani, Simone Callegari, Mia Palmkvist, Claudia Di Guglielmo & Maria G. Masucci

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Contributions

S.G. and M.G.M. designed the experiments; O.F. produced the recombinant lentiviruses; S.H., M.P and C.D.G. performed analysis; S.C. performed bioinformatics analysis; S.G. and M.G.M. wrote the manuscript.

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Correspondence to Maria G. Masucci.

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Gastaldello, S., Hildebrand, S., Faridani, O. et al. A deneddylase encoded by Epstein–Barr virus promotes viral DNA replication by regulating the activity of cullin-RING ligases. Nat Cell Biol 12, 351–361 (2010). https://doi.org/10.1038/ncb2035

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