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:	Nodoka Kasajima, Keita Matsuno, Hiroko Miyamoto, Masahiro Kajihara, Manabu Igarashi, Ayato Takada
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Ebola virus VP35 polymerase cofactor interferon antagonist IFN-inhibitory domain hydrophobic patch Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a0527a5bb6704eb7a1e2b8a7c96745d7
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spelling	oai:doaj.org-article:a0527a5bb6704eb7a1e2b8a7c96745d72021-11-25T19:14:31ZFunctional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain10.3390/v131123161999-4915https://doaj.org/article/a0527a5bb6704eb7a1e2b8a7c96745d72021-11-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2316https://doaj.org/toc/1999-4915Viral 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.Nodoka KasajimaKeita MatsunoHiroko MiyamotoMasahiro KajiharaManabu IgarashiAyato TakadaMDPI AGarticleEbola virusVP35polymerase cofactorinterferon antagonistIFN-inhibitory domainhydrophobic patchMicrobiologyQR1-502ENViruses, Vol 13, Iss 2316, p 2316 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ebola virus VP35 polymerase cofactor interferon antagonist IFN-inhibitory domain hydrophobic patch Microbiology QR1-502
spellingShingle	Ebola virus VP35 polymerase cofactor interferon antagonist IFN-inhibitory domain hydrophobic patch Microbiology QR1-502 Nodoka Kasajima Keita Matsuno Hiroko Miyamoto Masahiro Kajihara Manabu Igarashi Ayato Takada Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain
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.
format	article
author	Nodoka Kasajima Keita Matsuno Hiroko Miyamoto Masahiro Kajihara Manabu Igarashi Ayato Takada
author_facet	Nodoka Kasajima Keita Matsuno Hiroko Miyamoto Masahiro Kajihara Manabu Igarashi Ayato Takada
author_sort	Nodoka Kasajima
title	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_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_sort	functional importance of hydrophobic patches on the ebola virus vp35 ifn-inhibitory domain
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a0527a5bb6704eb7a1e2b8a7c96745d7
work_keys_str_mv	AT nodokakasajima functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain AT keitamatsuno functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain AT hirokomiyamoto functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain AT masahirokajihara functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain AT manabuigarashi functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain AT ayatotakada functionalimportanceofhydrophobicpatchesontheebolavirusvp35ifninhibitorydomain
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Functional Importance of Hydrophobic Patches on the Ebola Virus VP35 IFN-Inhibitory Domain

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