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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2022,7,26]]},"abstract":"<jats:p>Completion of the Lassa virus (LASV) life cycle critically depends on the activities of the virally encoded, RNA-dependent RNA polymerase in replication and transcription of the viral RNA genome in the cytoplasm of infected cells. The contribution of cellular proteins to these processes remains unclear. Here, we applied proximity proteomics to define the interactome of LASV polymerase in cells under conditions that recreate LASV RNA synthesis. We engineered a LASV polymerase-biotin ligase (TurboID) fusion protein that retained polymerase activity and successfully biotinylated the proximal proteome, which allowed the identification of 42 high-confidence LASV polymerase interactors. We subsequently performed a small interfering RNA (siRNA) screen to identify those interactors that have functional roles in authentic LASV infection. As proof of principle, we characterized eukaryotic peptide chain release factor subunit 3a (eRF3a\/GSPT1), which we found to be a proviral factor that physically associates with LASV polymerase. Targeted degradation of GSPT1 by a small-molecule drug candidate, CC-90009, resulted in strong inhibition of LASV infection in cultured cells. Our work demonstrates the feasibility of using proximity proteomics to illuminate and characterize yet-to-be-defined host-pathogen interactome, which can reveal new biology and uncover novel targets for the development of antivirals against highly pathogenic RNA viruses.<\/jats:p>","DOI":"10.1073\/pnas.2201208119","type":"journal-article","created":{"date-parts":[[2022,7,18]],"date-time":"2022-07-18T15:00:31Z","timestamp":1658156431000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":25,"title":["Proximity interactome analysis of Lassa polymerase reveals eRF3a\/GSPT1 as a druggable target for host-directed antivirals"],"prefix":"10.1073","volume":"119","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2482-2338","authenticated-orcid":false,"given":"Jingru","family":"Fang","sequence":"first","affiliation":[{"name":"Department of Immunology and Microbiology, Scripps Research, La Jolla, CA 92037"},{"name":"La Jolla Institute for Immunology, La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0759-3983","authenticated-orcid":false,"given":"Colette","family":"Pietzsch","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Texas Medical Branch, Galveston, TX 77550"},{"name":"Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX 77550"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0174-8454","authenticated-orcid":false,"given":"Haydar","family":"Witwit","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, Scripps Research, La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Tsaprailis","sequence":"additional","affiliation":[{"name":"Proteomics Core, Scripps Research, Jupiter, FL 33458"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6066-9900","authenticated-orcid":false,"given":"Gogce","family":"Crynen","sequence":"additional","affiliation":[{"name":"Bioinformatics and Statistics Core, Scripps Research, Jupiter, FL 33458"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelvin Frank","family":"Cho","sequence":"additional","affiliation":[{"name":"Cancer Biology Program, Stanford University, Stanford, CA 94305"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice Y.","family":"Ting","sequence":"additional","affiliation":[{"name":"Department of Genetics, Stanford University, Stanford, CA 94305"},{"name":"Department of Biology, Stanford University, Stanford, CA 94305"},{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305"},{"name":"Chan Zuckerberg Biohub, San Francisco, CA 94158"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0342-4824","authenticated-orcid":false,"given":"Alexander","family":"Bukreyev","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Texas Medical Branch, Galveston, TX 77550"},{"name":"Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX 77550"},{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77550"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1206-7451","authenticated-orcid":false,"given":"Erica Ollmann","family":"Saphire","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Immunology, La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8171-8115","authenticated-orcid":false,"given":"Juan Carlos","family":"de la Torre","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, Scripps Research, La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2022,7,18]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1080\/10408360490497456"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.idc.2019.08.002"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2502.180836"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2007.01113.x"},{"key":"e_1_3_4_5_2","first-page":"231","article-title":"Arenavirus quasispecies and their biological implications","volume":"392","author":"Grande-P\u00e9rez A.","year":"2016","unstructured":"A. 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