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The VP24 protein encoded by members of the Ebolavirus genus blocks type I interferon (IFN-I) signaling through interaction with host karyopherin alpha nuclear transporters, potentially contributing to virulence. Previously, we demonstrated that BDBV VP24 (bVP24) binds with lower affinities to karyopherin alpha proteins relative to EBOV VP24 (eVP24), and this correlated with a reduced inhibition in IFN-I signaling. We hypothesized that modification of eVP24-karyopherin alpha interface to make it similar to bVP24 would attenuate the ability to antagonize IFN-I response. We generated a panel of recombinant EBOVs containing single or combinations of point mutations in the eVP24-karyopherin alpha interface. Most of the viruses appeared to be attenuated in both IFN-I-competent 769-P and IFN-I-deficient Vero-E6 cells in the presence of IFNs. However, the R140A mutant grew at reduced levels even in the absence of IFNs in both cell lines, as well as in U3A STAT1 knockout cells. Both the R140A mutation and its combination with the N135A mutation greatly reduced the amounts of viral genomic RNA and mRNA suggesting that these mutations attenuate the virus in an IFN-I-independent attenuation. Additionally, we found that unlike eVP24, bVP24 does not inhibit interferon lambda 1 (IFN-\u03bb1), interferon beta (IFN-\u03b2), and ISG15, which potentially explains the lower pathogenicity of BDBV relative to EBOV. Thus, the VP24 residues binding karyopherin alpha attenuates the virus by IFN-I-dependent and independent mechanisms.<\/jats:p>","DOI":"10.3390\/v15051075","type":"journal-article","created":{"date-parts":[[2023,4,28]],"date-time":"2023-04-28T04:36:15Z","timestamp":1682656575000},"page":"1075","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Ebolavirus Species-Specific Interferon Antagonism Mediated by VP24"],"prefix":"10.3390","volume":"15","author":[{"given":"Palaniappan","family":"Ramanathan","sequence":"first","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bersabeh","family":"Tigabu","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo I.","family":"Santos","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philipp A.","family":"Ilinykh","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Kuzmina","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivia A.","family":"Vogel","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naveen","family":"Thakur","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamza","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9342-1968","authenticated-orcid":false,"given":"Chao","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaya K.","family":"Amarasinghe","sequence":"additional","affiliation":[{"name":"Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4195-425X","authenticated-orcid":false,"given":"Christopher F.","family":"Basler","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Bukreyev","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Galveston National Laboratory, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"},{"name":"Department of Microbiology & Immunology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1084","DOI":"10.1038\/s41564-018-0227-2","article-title":"The discovery of Bombali virus adds further support for bats as hosts of ebolaviruses","volume":"3","author":"Goldstein","year":"2018","journal-title":"Nat. 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