{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T20:34:02Z","timestamp":1768682042744,"version":"3.49.0"},"reference-count":36,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,20]],"date-time":"2021-11-20T00:00:00Z","timestamp":1637366400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["20H03140"],"award-info":[{"award-number":["20H03140"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP20fm0208101"],"award-info":[{"award-number":["JP20fm0208101"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP19fm0208001"],"award-info":[{"award-number":["JP19fm0208001"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Viral protein 35 (VP35) of Ebola virus (EBOV) is a multifunctional protein that mainly acts as a viral polymerase cofactor and an interferon antagonist. VP35 interacts with the viral nucleoprotein (NP) and double-stranded RNA for viral RNA transcription\/replication and inhibition of type I interferon (IFN) production, respectively. The C-terminal portion of VP35, which is termed the IFN-inhibitory domain (IID), is important for both functions. To further identify critical regions in this domain, we analyzed the physical properties of the surface of VP35 IID, focusing on hydrophobic patches, which are expected to be functional sites that are involved in interactions with other molecules. Based on the known structural information of VP35 IID, three hydrophobic patches were identified on its surface and their biological importance was investigated using minigenome and IFN-\u03b2 promoter-reporter assays. Site-directed mutagenesis revealed that some of the amino acid substitutions that were predicted to disrupt the hydrophobicity of the patches significantly decreased the efficiency of viral genome replication\/transcription due to reduced interaction with NP, suggesting that the hydrophobic patches might be critical for the formation of a replication complex through the interaction with NP. It was also found that the hydrophobic patches were involved in the IFN-inhibitory function of VP35. These results highlight the importance of hydrophobic patches on the surface of EBOV VP35 IID and also indicate that patch analysis is useful for the identification of amino acid residues that directly contribute to protein functions.<\/jats:p>","DOI":"10.3390\/v13112316","type":"journal-article","created":{"date-parts":[[2021,11,21]],"date-time":"2021-11-21T21:02:06Z","timestamp":1637528526000},"page":"2316","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain"],"prefix":"10.3390","volume":"13","author":[{"given":"Nodoka","family":"Kasajima","sequence":"first","affiliation":[{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4205-6526","authenticated-orcid":false,"given":"Keita","family":"Matsuno","sequence":"additional","affiliation":[{"name":"Division of Risk Analysis and Management, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"One Health Research Center, Hokkaido University, Sapporo 001-0020, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroko","family":"Miyamoto","sequence":"additional","affiliation":[{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Kajihara","sequence":"additional","affiliation":[{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5625-1357","authenticated-orcid":false,"given":"Manabu","family":"Igarashi","sequence":"additional","affiliation":[{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education, Hokkaido University, Sapporo 001-0020, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2464-6642","authenticated-orcid":false,"given":"Ayato","family":"Takada","sequence":"additional","affiliation":[{"name":"Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"One Health Research Center, Hokkaido University, Sapporo 001-0020, Japan"},{"name":"School of Veterinary Medicine, The University of Zambia, Lusaka P.O. Box 32379, Zambia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41572-020-0147-3","article-title":"Ebola virus disease","volume":"6","author":"Jacob","year":"2020","journal-title":"Nat. Rev. Dis. Primers."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2283","DOI":"10.1007\/s00705-018-3814-x","article-title":"Taxonomy of the order Mononegavirales: Update 2018","volume":"163","author":"Amarasinghe","year":"2018","journal-title":"Arch. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1038\/nrd.2017.251","article-title":"Post-exposure treatments for Ebola and Marburg virus infections","volume":"17","author":"Cross","year":"2018","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.ppat.1002304","article-title":"Discovery of an ebolavirus-like filovirus in europe","volume":"7","author":"Negredo","year":"2011","journal-title":"PLoS Pathog."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1038\/s41564-018-0328-y","article-title":"Characterization of a filovirus (M\u011bngl\u00e0 virus) from Rousettus bats in China","volume":"4","author":"Yang","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1038\/nrmicro3524","article-title":"Filovirus pathogenesis and immune evasion: Insights from Ebola virus and Marburg virus","volume":"13","author":"Messaoudi","year":"2015","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_7","first-page":"1","article-title":"Ebolavirus interferon antagonists\u2014protein interaction perspectives to combat pathogenesis","volume":"17","author":"Banerjee","year":"2017","journal-title":"Brief. Funct. Genom."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3069","DOI":"10.1128\/JVI.01875-08","article-title":"Ebola virus protein VP35 impairs the function of interferon regulatory factor-activating kinases IKK and TBK-1","volume":"83","author":"Prins","year":"2009","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1089\/jir.2009.0076","article-title":"Evasion of interferon responses by Ebola and Marburg viruses","volume":"29","author":"Basler","year":"2009","journal-title":"J. Interf. Cytokine Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.virol.2005.06.044","article-title":"Homo-oligomerization facilitates the interferon-antagonist activity of the ebolavirus VP35 protein","volume":"341","author":"Reid","year":"2005","journal-title":"Virology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7945","DOI":"10.1128\/JVI.77.14.7945-7956.2003","article-title":"The Ebola virus VP35 protein inhibits activation of interferon regulatory factor 3","volume":"77","author":"Basler","year":"2003","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5168","DOI":"10.1128\/JVI.02199-05","article-title":"Ebola virus VP35 protein binds double-stranded RNA and inhibits alpha\/beta interferon production induced by RIG-I signaling","volume":"80","author":"Cardenas","year":"2006","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.virol.2004.07.006","article-title":"A C-terminal basic amino acid motif of Zaire ebolavirus VP35 is essential for type I interferon antagonism and displays high identity with the RNA-binding domain of another interferon antagonist, the NS1 protein of influenza a virus","volume":"328","author":"Hartman","year":"2004","journal-title":"Virology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1073\/pnas.0807854106","article-title":"Structure of the Ebola VP35 interferon inhibitory domain","volume":"106","author":"Leung","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"10581","DOI":"10.1128\/JVI.00925-10","article-title":"Basic residues within the ebolavirus VP35 protein are required for its viral polymerase cofactor function","volume":"84","author":"Prins","year":"2010","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1038\/nsmb.1765","article-title":"Structural basis for dsRNA recognition and interferon antagonism by Ebola VP35","volume":"17","author":"Leung","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3004","DOI":"10.1128\/JVI.02459-09","article-title":"Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in Guinea Pigs","volume":"84","author":"Prins","year":"2010","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2699","DOI":"10.1128\/JVI.02344-07","article-title":"Inhibition of IRF-3 activation by VP35 is critical for the high level of virulence of Ebola virus","volume":"82","author":"Hartman","year":"2008","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1006\/jmbi.1997.1234","article-title":"Analysis of protein-protein interaction sites using surface patches","volume":"272","author":"Jones","year":"1997","journal-title":"J. Mol. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1006\/jmbi.1997.1233","article-title":"Prediction of protein-protein interaction sites using patch analysis","volume":"272","author":"Jones","year":"1997","journal-title":"J. Mol. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/0378-1119(91)90434-D","article-title":"Efficient secletion for high-expression transformants with a novel eukaryotic vector","volume":"108","author":"Niwa","year":"1991","journal-title":"Gene"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kondoh, T., Manzoor, R., Nao, N., Maruyama, J., Furuyama, W., Miyamoto, H., Shigeno, A., Kuroda, M., Matsuno, K., and Fujikura, D. (2017). Putative endogenous filovirus VP35-like protein potentially functions as an IFN antagonist but not a polymerase cofactor. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0186450"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"S284","DOI":"10.1086\/520582","article-title":"Ebola virus (EBOV) VP24 inhibits transcription and replication of the EBOV genome","volume":"196","author":"Watanabe","year":"2007","journal-title":"J. Infect. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1128\/JVI.78.2.999-1005.2004","article-title":"Production of novel Ebola virus-like particles from cDNAs: An alternative to Ebola virus generation by reverse genetics","volume":"78","author":"Watanabe","year":"2004","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.virusres.2013.05.004","article-title":"Mapping of conserved and species-specific antibody epitopes on the Ebola virus nucleoprotein","volume":"176","author":"Changula","year":"2013","journal-title":"Virus Res."},{"key":"ref_26","unstructured":"Team, R.C. (2021, September 15). R: A Language and Environment for Statistical Computing. Available online: https:\/\/www.r-project.org\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1073\/pnas.0910547107","article-title":"Ebolavirus VP35 uses a bimodal strategy to bind dsRNA for innate immune suppression","volume":"107","author":"Kimberlin","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"0794","DOI":"10.1371\/journal.ppat.0030086","article-title":"The ebola virus VP35 protein is a suppressor of RNA silencing","volume":"3","author":"Haasnoot","year":"2007","journal-title":"PLoS Pathog."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1006\/viro.1998.9328","article-title":"Interactions of Marburg virus nucleocapsid proteins","volume":"249","author":"Becker","year":"1998","journal-title":"Virology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1128\/JVI.73.3.2333-2342.1999","article-title":"Comparison of the transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems","volume":"73","author":"Muhlberger","year":"1999","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"65","DOI":"10.21660\/2018.49.3535","article-title":"Fragment-based lead compound design to inhibit Ebola VP35 through computational studies","volume":"15","author":"Marnolia","year":"2018","journal-title":"Int. J. Geomate"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s10867-016-9440-5","article-title":"Molecular dynamics exploration of the binding mechanism and properties of single-walled carbon nanotube to WT and mutant VP35 FBP region of Ebola virus","volume":"43","author":"Zhang","year":"2017","journal-title":"J. Biol. Phys."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1016\/j.jmb.2014.01.010","article-title":"In silico derived small molecules bind the filovirus VP35 protein and inhibit its polymerase co-factor activity","volume":"426","author":"Brown","year":"2014","journal-title":"J. Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"20661","DOI":"10.1073\/pnas.1213559109","article-title":"Structural basis for Marburg virus VP35-mediated immune evasion mechanisms","volume":"109","author":"Ramanan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.virol.2015.07.010","article-title":"Lloviu virus VP24 and VP35 proteins function as innate immune antagonists in human and bat cells","volume":"485","author":"Feagins","year":"2015","journal-title":"Virology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e00191-20","DOI":"10.1128\/JVI.00191-20","article-title":"Impact of M\u011bngl\u00e0 virus proteins on human and bat innate immune pathways","volume":"94","author":"Williams","year":"2020","journal-title":"J. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/11\/2316\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:33:12Z","timestamp":1760167992000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/11\/2316"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,20]]},"references-count":36,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["v13112316"],"URL":"https:\/\/doi.org\/10.3390\/v13112316","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,20]]}}}