The Role of Apoptosis in the Biology of New World Arenaviruses
Final Report Abstract
Arenaviruses include important agents of severe human disease, including haemorrhagic fever. However, these viruses also exhibit marked diversity in their pathogenicity, with highly pathogenic (e.g. Junín virus, JUNV) and apathogenic viruses (e.g. Tacaribe virus, TCRV) often being closely related. Among the factors that appear to correlate with this phenomenon is a difference in their ability to regulate apoptosis. It was, therefore, the goal of this project to better understand the mechanisms by which apoptosis is being regulated by JUNV and TCRV, and its biological relevance during virus infection. The initial project phase showed that both TCRV and JUNV infections trigger the intrinsic apoptotic pathway through upregulation of the pro-apoptotic BH3-only proteins Puma and Noxa, as a result of activation of their transcription factor p53, and that in the case of TCRV, this process involves phosphorylation of p53 at Ser392. At the same time, these signals were also moderated by anti-apoptotic changes in Bad, including phosphorylation at position Ser112. However, in contrast to TCRV, where apoptotic cell death occurs in response to these signalling events, we could demonstrate that for JUNV, this process is impaired by cleavage of the JUNV nucleoprotein (NP) as a decoy substrate for activated caspases. A similar cleavage is not possible for TCRV NP, and our data suggest this is due to its inability to generate a large soluble NP isoform (NP53kD), which we have now shown is produced by alternative in-frame translation. Finally, we could generate JUNV mutants that were unable to produce NP isoforms and thereby definitively establish their role in preventing apoptotic cell death during JUNV infection. Since our results indicated that arenavirus-induced apoptosis is being regulated by phosphorylation events, the goal of the follow-up phase was to identify the kinases involved and to better understand their role in virus infection. Here we could demonstrate robust activation of both p38 and JNK in response to infection with either TCRV and JUNV, consistent with their role in p53 activation. Further, we found that inhibition of either JNK or p38 markedly impaired viral replication, although inhibition of apoptotic cell death itself does not. An interactomics approach further revealed interaction of Z, which we found to be responsible for triggering apoptosis, with the upstream kinase ASK1, suggesting that this interaction is the mechanism leading to p38 and JNK activation. Surprisingly, despite known functions of p38 and JNK in cytokine regulation, we did not observe notable differences in the host response to infection with JUNV and TCRV. Taken together our work on these projects has allowed us establish a model by which arenavirus infections induce the apoptotic signalling, and also differentially regulate these pathways through NP cleavage, thereby resulting in the production of truncated isoforms of JUNV NP with altered localization. Further, we have shown that activation of the kinases involved in the apoptotic signalling cascade are essential for efficient virus replication, and as such may present viable antiviral targets.
Publications
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Deciphering apoptotic host cell responses activated by Tacaribe virus infection. 17th Workshop "Cell Biology of Viral Infections" of the Society for Virology (GfV) 2018, oral presentation
Holzerland J. & Groseth A.
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Functional analysis of Junín nucleoprotein variants for their role in the viral life cycle. 17th Workshop "Cell Biology of Viral Infections" of the Society for Virology (GfV) 2018, oral presentation
Bostedt L. & Groseth A.
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Assessing cross-reactivity of Junín virus-directed neutralizing antibodies. Antiviral Research, 163, 106-116.
Leske, Anne; Waßmann, Irke; Schnepel, Kevin; Shifflett, Kyle; Holzerland, Julia; Bostedt, Linus; Bohn, Patrick; Mettenleiter, Thomas C.; Briggiler, Ana M.; Brignone, Julia; Enria, Delia; Cordo, Sandra M.; Hoenen, Thomas & Groseth, Allison
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Bad, Noxa and Puma are Key Regulators of Tacaribe virus-induced Apoptosis. 18th Workshop "Cell Biology of Viral Infections" of the Society for Virology (GfV) 2019, oral presentation
Holzerland J. & Groseth A.
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High-throughput screening for negative-stranded hemorrhagic fever viruses using reverse genetics. Antiviral Research, 170, 104569.
Wendt, Lisa; Bostedt, Linus; Hoenen, Thomas & Groseth, Allison
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Junín Virus Nucleoprotein Isoforms and their Functional Role in the Viral Infection Cycle. 29th Annual Meeting of the Society for Virology (GfV) 2019, poster presentation
Bostedt L. & Groseth A.
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Tacaribe virus induces classical intrinsic apoptosis by activating BH3-only proteins. 29th Annual Meeting of the Society for Virology (GfV) 2019, poster presentation
Holzerland J. & Groseth A.
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Unravelling Junín Virus Nucleoprotein Isoforms Activities in the Viral Life Cycle. 18th Workshop "Cell Biology of Viral Infections" of the Society for Virology (GfV) 2019, oral presentation
Bostedt L. & Groseth A.
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BH3-only sensors Bad, Noxa and Puma are Key Regulators of Tacaribe virus-induced Apoptosis. PLOS Pathogens, 16(10), e1008948.
Holzerland, Julia; Fénéant, Lucie; Banadyga, Logan; Hölper, Julia E.; Knittler, Michael R. & Groseth, Allison
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Complete genome sequence of Tacaribe virus. Archives of Virology, 165(8), 1899-1903.
Holzerland, Julia; Leske, Anne; Fénéant, Lucie; Garcin, Dominique; Kolakofsky, Daniel & Groseth, Allison
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BH3-only sensors Bad, Noxa and Puma are Key Regulators of Arenavirus-induced Apoptosis. 30th Annual Meeting of the Society for Virology (GfV) 2021, oral presentation
Holzerland J., Fénéant L., Banadyga L., Hölper J.E., Knittler M.R. & Groseth, A.
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BH3-only sensors Bad, Noxa and Puma are Key Regulators of Arenavirus-induced Apoptosis. 40th Annual Meeting of the American Society for Virology 2021, oral presentation
Groseth A., Fénéant L. & Holzerland J.
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Identification of host cell signaling molecules responsible for apoptosis regulation in response to arenavirus infection. 19th Workshop "Cell Biology of Viral Infections" of the Society for Virology (GfV) 2021, oral presentation
Holzerland J., Fénéant, F. & Groseth A.
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Connecting Host Cell Response, Apoptosis and Arenavirus Pathogenesis. 18th NSV meeting 2022, oral presentation
Holzerland J., Fénéant L., Bostedt L. & Groseth A.
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Regulation of host cell signaling molecules in response to arenavirus infection. 31st Annual Meeting of the Society for Virology (GfV) 2022, poster presentation
Holzerland J., Fénéant L. & Groseth, A.
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Regulation of Stress-Activated Kinases in Response to Tacaribe Virus Infection and Its Implications for Viral Replication. Viruses, 14(9), 2018.
Holzerland, Julia; Fénéant, Lucie & Groseth, Allison
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The Arenavirus-Host Cell Interface: at the crossroads of virus replication, host cell response and pathogenesis. LabRoots 9th Annual Microbiology Week 2023, invited speaker
Groseth A.
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Understanding the Arenavirus-Host Cell Interface as a Guide to the Development of Novel Antiviral Approaches. 36th International Conference on Antiviral Research (ICAR) 2023, invited speaker
Groseth A.
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Unravelling the Host Cell Response to Arenavirus Infection. 2nd Workshop " One Health and Zoonotic Viruses " of the Society for Virology (GfV) 2023, keynote lecture
Groseth A.
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Alternative translation contributes to the generation of a cytoplasmic subpopulation of the Junín virus nucleoprotein that inhibits caspase activation and innate immunity. Journal of Virology, 98(2).
Bostedt, Linus; Fénéant, Lucie; Leske, Anne; Holzerland, Julia; Günther, Karla; Waßmann, Irke; Bohn, Patrick & Groseth, Allison
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Exploring the arenavirus-host interface as a path to novel antiviral approaches. 8th National Congress of the Italian Society for Virology (ISV) 2024, invited speaker
Groseth A.
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Virus-Host Interactions: Connecting Mechanisms with Implications for Arenavirus Pathogenesis. Bunya 2024, invited speaker
Groseth, A.
