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Oncolytic reovirus induces intracellular redistribution of Ras to promote apoptosis and progeny virus release

Oncogene volume 35, pages 771–782 (2016)Cite this article

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Abstract

Reovirus is a naturally oncolytic virus that preferentially replicates in Ras-transformed cells and is currently undergoing clinical trials as a cancer therapeutic. Ras transformation promotes reovirus oncolysis by enhancing virion disassembly during entry, viral progeny production, and virus release through apoptosis; however, the mechanism behind the latter is not well understood. Here, we show that reovirus alters the intracellular location of oncogenic Ras to induce apoptosis of H-RasV12-transformed fibroblasts. Reovirus infection decreases Ras palmitoylation levels and causes accumulation of Ras in the Golgi through Golgi fragmentation. With the Golgi being the site of Ras palmitoylation, treatment of target cells with the palmitoylation inhibitor, 2-bromopalmitate (2BP), prompts a greater accumulation of H-RasV12 in the Golgi, and a dose-dependent increase in progeny virus release and subsequent spread. Conversely, tethering H-RasV12 to the plasma membrane (thereby preventing its movement to the Golgi) allows for efficient virus production, but results in basal levels of reovirus-induced cell death. Analysis of Ras downstream signaling reveals that cells expressing cycling H-RasV12 have elevated levels of phosphorylated JNK (c-Jun N-terminal kinase), and that Ras retained at the Golgi body by 2BP increases activation of the MEKK1/MKK4/JNK signaling pathway to promote cell death. Collectively, our data suggest that reovirus induces Golgi fragmentation of target cells, and the subsequent accumulation of oncogenic Ras in the Golgi body initiates apoptotic signaling events required for virus release and spread.

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Acknowledgements

This work was supported by the research grants from the Canadian Institutes for Health Research (Grant FRN # 130574) and the Terry Fox Research Institute (TFRI) New Frontiers Program in Cancer Research (through Canadian Oncolytic Virus Consortium (COVCo)) to PWKL and SAG. Funding for KA Garant was awarded through the National Sciences and Engineering Research Council of Canada, the Killam Trusts and the Nova Scotia Health Research Foundation. Plasma membrane-tethered CD8-H-RasV12xx in pCEFL was a gift from Dr Piero Crespo (University of Cantabria, Cantabria, Spain). D-G Ahn was supported by the Canadian Institute of Health Research (CIHR)—Cancer Research Training Program and the Beatrice Hunter Cancer Research Institute (BHCRI). SG was supported by CIHR postdoctoral fellowship.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Dalhousie University, Halifax, NS, Canada

    K A Garant, D-G Ahn, S A Gujar & P W K Lee

  2. Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB, Canada

    M Shmulevitz

  3. Division of Fundamental Neurobiology, Toronto Western Research Institute, Toronto, ON, Canada

    L Pan

  4. Department of Oceanography, Dalhousie University, Halifax, NS, Canada

    R M Daigle

  5. Strategy and Organizational Performance, IWK Health Centre, Halifax, NS, Canada

    S A Gujar

  6. Department of Pathology, Dalhousie University, Halifax, NS, Canada

    P W K Lee

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Correspondence to S A Gujar or P W K Lee.

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Garant, K., Shmulevitz, M., Pan, L. et al. Oncolytic reovirus induces intracellular redistribution of Ras to promote apoptosis and progeny virus release. Oncogene 35, 771–782 (2016). https://doi.org/10.1038/onc.2015.136

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