Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment

ABSTRACT The filoviruses are etiological agents of life-threatening hemorrhagic fever with high mortality rate and risk of potential outbreak. Among members of this family, the Ebola (EBOV), Sudan (SUDV), and Marburg (MARV) viruses are considered the most pathogenic for humans. The ebolavirus nucleo...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sara Landeras-Bueno, Shun-ichiro Oda, Michael J. Norris, Zhe Li Salie, Javier Guenaga, Richard T. Wyatt, Erica Ollmann Saphire
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	RNA virus replication Sudan virus crystal structure ebolavirus nucleoprotein phosphoprotein Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/e8b10b7a660545c983dc47c5094b4419
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:e8b10b7a660545c983dc47c5094b4419
record_format	dspace
spelling	oai:doaj.org-article:e8b10b7a660545c983dc47c5094b44192021-11-15T16:22:10ZSudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment10.1128/mBio.00734-192150-7511https://doaj.org/article/e8b10b7a660545c983dc47c5094b44192019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00734-19https://doaj.org/toc/2150-7511ABSTRACT The filoviruses are etiological agents of life-threatening hemorrhagic fever with high mortality rate and risk of potential outbreak. Among members of this family, the Ebola (EBOV), Sudan (SUDV), and Marburg (MARV) viruses are considered the most pathogenic for humans. The ebolavirus nucleoprotein (NP) is the most abundant protein in infected cells and is essential for viral transcription and replication; thus, it represents an attractive target for therapeutic intervention. Here, we present the structure of SUDV NP in complex with the amino-terminal portion of the phosphoprotein VP35 at 2.3 Å. This structure captures VP35 chaperoning SUDV NP in a monomeric and RNA-free state. This transient state has been proposed to be key to maintaining a pool of monomeric and RNA-free NPs prior to NP-NP polymerization and encapsidation of the viral RNA genome. This structure also reveals a newly visualized interaction between NP and VP35, a well-defined beta sheet that is not present in previous structures. Affinity binding assays demonstrate that this beta sheet is essential for maintaining the high-affinity interaction between VP35 and a hydrophobic pocket on SUDV NP, and electron microscopy indicates the importance of this binding interaction to the oligomeric state and assembly of NP in human cells. Complementary structure-directed mutagenesis identifies critical residues conserved across the filovirus family that could be targeted by broadly effective antivirals. IMPORTANCE Outbreaks of the filoviruses can be unpredictable in timing, location, and identity of the causative virus, with each of Ebola virus, Sudan virus, Bundibugyo virus, and Marburg virus reemerging in the last several years to cause human disease with 30 to 90% lethality. The 2014–2016 outbreak in particular, with nearly 30,000 patients, highlighted the ability of these viruses to emerge unexpectedly and spread rapidly. Two ebolavirus outbreaks have emerged this year, yet we still lack FDA-approved drugs with pan-filovirus activity to treat existing and emergent ebolaviruses. For all filoviruses, the interaction between the nucleoprotein and the phosphoprotein is essential for the virus life cycle and is a potential target for therapeutic intervention. In this report, we describe the crystal structure of the SUDV nucleoprotein with the interacting domain of the viral phosphoprotein, and we identify residues critical for high-affinity interaction and for control of the oligomeric state of the nucleoprotein. Structural comparison of this heterodimer with other members of the filovirus family allowed us to find conserved and essential atomic features that will facilitate understanding of the virus life cycle and the rational design of antivirals.Sara Landeras-BuenoShun-ichiro OdaMichael J. NorrisZhe Li SalieJavier GuenagaRichard T. WyattErica Ollmann SaphireAmerican Society for MicrobiologyarticleRNA virus replicationSudan viruscrystal structureebolavirusnucleoproteinphosphoproteinMicrobiologyQR1-502ENmBio, Vol 10, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	RNA virus replication Sudan virus crystal structure ebolavirus nucleoprotein phosphoprotein Microbiology QR1-502
spellingShingle	RNA virus replication Sudan virus crystal structure ebolavirus nucleoprotein phosphoprotein Microbiology QR1-502 Sara Landeras-Bueno Shun-ichiro Oda Michael J. Norris Zhe Li Salie Javier Guenaga Richard T. Wyatt Erica Ollmann Saphire Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
description	ABSTRACT The filoviruses are etiological agents of life-threatening hemorrhagic fever with high mortality rate and risk of potential outbreak. Among members of this family, the Ebola (EBOV), Sudan (SUDV), and Marburg (MARV) viruses are considered the most pathogenic for humans. The ebolavirus nucleoprotein (NP) is the most abundant protein in infected cells and is essential for viral transcription and replication; thus, it represents an attractive target for therapeutic intervention. Here, we present the structure of SUDV NP in complex with the amino-terminal portion of the phosphoprotein VP35 at 2.3 Å. This structure captures VP35 chaperoning SUDV NP in a monomeric and RNA-free state. This transient state has been proposed to be key to maintaining a pool of monomeric and RNA-free NPs prior to NP-NP polymerization and encapsidation of the viral RNA genome. This structure also reveals a newly visualized interaction between NP and VP35, a well-defined beta sheet that is not present in previous structures. Affinity binding assays demonstrate that this beta sheet is essential for maintaining the high-affinity interaction between VP35 and a hydrophobic pocket on SUDV NP, and electron microscopy indicates the importance of this binding interaction to the oligomeric state and assembly of NP in human cells. Complementary structure-directed mutagenesis identifies critical residues conserved across the filovirus family that could be targeted by broadly effective antivirals. IMPORTANCE Outbreaks of the filoviruses can be unpredictable in timing, location, and identity of the causative virus, with each of Ebola virus, Sudan virus, Bundibugyo virus, and Marburg virus reemerging in the last several years to cause human disease with 30 to 90% lethality. The 2014–2016 outbreak in particular, with nearly 30,000 patients, highlighted the ability of these viruses to emerge unexpectedly and spread rapidly. Two ebolavirus outbreaks have emerged this year, yet we still lack FDA-approved drugs with pan-filovirus activity to treat existing and emergent ebolaviruses. For all filoviruses, the interaction between the nucleoprotein and the phosphoprotein is essential for the virus life cycle and is a potential target for therapeutic intervention. In this report, we describe the crystal structure of the SUDV nucleoprotein with the interacting domain of the viral phosphoprotein, and we identify residues critical for high-affinity interaction and for control of the oligomeric state of the nucleoprotein. Structural comparison of this heterodimer with other members of the filovirus family allowed us to find conserved and essential atomic features that will facilitate understanding of the virus life cycle and the rational design of antivirals.
format	article
author	Sara Landeras-Bueno Shun-ichiro Oda Michael J. Norris Zhe Li Salie Javier Guenaga Richard T. Wyatt Erica Ollmann Saphire
author_facet	Sara Landeras-Bueno Shun-ichiro Oda Michael J. Norris Zhe Li Salie Javier Guenaga Richard T. Wyatt Erica Ollmann Saphire
author_sort	Sara Landeras-Bueno
title	Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
title_short	Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
title_full	Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
title_fullStr	Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
title_full_unstemmed	Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment
title_sort	sudan ebolavirus vp35-np crystal structure reveals a potential target for pan-filovirus treatment
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/e8b10b7a660545c983dc47c5094b4419
work_keys_str_mv	AT saralanderasbueno sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT shunichirooda sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT michaeljnorris sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT zhelisalie sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT javierguenaga sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT richardtwyatt sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment AT ericaollmannsaphire sudanebolavirusvp35npcrystalstructurerevealsapotentialtargetforpanfilovirustreatment
_version_	1718426940669427712

Sudan Ebolavirus VP35-NP Crystal Structure Reveals a Potential Target for Pan-Filovirus Treatment

Ejemplares similares