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Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain

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 interfero...

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Autores principales: Kasajima, Nodoka, Matsuno, Keita, Miyamoto, Hiroko, Kajihara, Masahiro, Igarashi, Manabu, Takada, Ayato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8618116/
https://www.ncbi.nlm.nih.gov/pubmed/34835122
http://dx.doi.org/10.3390/v13112316
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author Kasajima, Nodoka
Matsuno, Keita
Miyamoto, Hiroko
Kajihara, Masahiro
Igarashi, Manabu
Takada, Ayato
author_facet Kasajima, Nodoka
Matsuno, Keita
Miyamoto, Hiroko
Kajihara, Masahiro
Igarashi, Manabu
Takada, Ayato
author_sort Kasajima, Nodoka
collection PubMed
description 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-β 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.
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spelling pubmed-86181162021-11-27 Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain Kasajima, Nodoka Matsuno, Keita Miyamoto, Hiroko Kajihara, Masahiro Igarashi, Manabu Takada, Ayato Viruses Article 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-β 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. MDPI 2021-11-20 /pmc/articles/PMC8618116/ /pubmed/34835122 http://dx.doi.org/10.3390/v13112316 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kasajima, Nodoka
Matsuno, Keita
Miyamoto, Hiroko
Kajihara, Masahiro
Igarashi, Manabu
Takada, Ayato
Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title_full Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title_fullStr Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title_full_unstemmed Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title_short Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
title_sort functional importance of hydrophobic patches on the ebola virus vp35 ifn-inhibitory domain
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8618116/
https://www.ncbi.nlm.nih.gov/pubmed/34835122
http://dx.doi.org/10.3390/v13112316
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