@article{f1275879957943ae918a8bcd2d864974,
title = "Functional interactomes of the Ebola virus polymerase identified by proximity proteomics in the context of viral replication",
abstract = "Ebola virus (EBOV) critically depends on the viral polymerase to replicate and transcribe the viral RNA genome in the cytoplasm of host cells, where cellular factors can antagonize or facilitate the virus life cycle. Here we leverage proximity proteomics and conduct a small interfering RNA (siRNA) screen to define the functional interactome of EBOV polymerase. As a proof of principle, we validate two cellular mRNA decay factors from 35 identified host factors: eukaryotic peptide chain release factor subunit 3a (eRF3a/GSPT1) and up-frameshift protein 1 (UPF1). Our data suggest that EBOV can subvert restrictions of cellular mRNA decay and repurpose GSPT1 and UPF1 to promote viral replication. Treating EBOV-infected human hepatocytes with a drug candidate that targets GSPT1 for degradation significantly reduces viral RNA load and particle production. Our work demonstrates the utility of proximity proteomics to capture the functional host interactome of the EBOV polymerase and to illuminate host-dependent regulation of viral RNA synthesis.",
keywords = "CP: Microbiology, Ebola virus, RNA-dependent RNA polymerase, RdRP, eukaryotic peptide chain release factor subunit 3a, eRF3a/GSPT1, host-directed antiviral targets, host-virus interaction, nonsense-mediated mRNA decay, NMD, proximity proteomics, viral RNA synthesis",
author = "Jingru Fang and Colette Pietzsch and George Tsaprailis and Gogce Crynen and Cho, {Kelvin Frank} and Ting, {Alice Y.} and Alexander Bukreyev and {de la Torre}, {Juan Carlos} and Saphire, {Erica Ollmann}",
note = "Funding Information: We thank Jonathan Towner (CDC) and Stuart Nichol (CDC) for providing the EBOV full-length clone and Yoshihiro Kawaoka (University of Wisconsin) for providing the pCEZ-NP, VP35, L, VP30, and pHH21-3E5E-Fluc plasmids and sharing the anti-VP30 antibody. We thank Beatrice Cubitt from the de la Torre Lab (Scripps Research, CA) for helping with cloning of the pCI-FLAG-GSPT1 plasmid. We thank Diptiben Parekh (LJI) for plasmid preparations, Sharon Schendel (LJI) for manuscript editing, Zbigniew Mikulski of the Microscopy Core Facility (LJI) for microscopy training, and NIH S10OD021831 for sponsoring the Zeiss LSM 880 microscope. This research was supported by institutional funds of University of Texas Medical Branch (A.B.), NIAID grants AI125626 and AI128556 (J.C.T.), and institutional funds of the La Jolla Institute for Immunology (E.O.S.). J.F. was supported by the Donald E. and Delia B. Baxter Foundation fellowship. J.F. J.C.T. and E.O.S. designed research and wrote the paper. J.F. performed all non-BSL4 experiments and analyzed data. C.P. performed all BSL-4 experiments. A.B. provided supervision for the BSL-4 research. G.T. performed proteomics experiments. G.C. performed the experimental design and data analysis for proteomics. K.F.C. and A.Y.T. provided a critical resource. All authors edited and approved the paper. The authors declare no competing interests. Funding Information: We thank Jonathan Towner (CDC) and Stuart Nichol (CDC) for providing the EBOV full-length clone and Yoshihiro Kawaoka (University of Wisconsin) for providing the pCEZ-NP, VP35, L, VP30, and pHH21-3E5E-Fluc plasmids and sharing the anti-VP30 antibody. We thank Beatrice Cubitt from the de la Torre Lab (Scripps Research, CA) for helping with cloning of the pCI-FLAG-GSPT1 plasmid. We thank Diptiben Parekh (LJI) for plasmid preparations, Sharon Schendel (LJI) for manuscript editing, Zbigniew Mikulski of the Microscopy Core Facility (LJI) for microscopy training, and NIH S10OD021831 for sponsoring the Zeiss LSM 880 microscope. This research was supported by institutional funds of University of Texas Medical Branch (A.B.), NIAID grants AI125626 and AI128556 (J.C.T.), and institutional funds of the La Jolla Institute for Immunology (E.O.S.). J.F. was supported by the Donald E. and Delia B. Baxter Foundation fellowship. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = mar,
day = "22",
doi = "10.1016/j.celrep.2022.110544",
language = "English (US)",
volume = "38",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "12",
}