The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing.
Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antivi...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2017-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0178717 |
| _version_ | 1818586714116980736 |
|---|---|
| author | Kari A Dilley Alexander A Voorhies Priya Luthra Vinita Puri Timothy B Stockwell Hernan Lorenzi Christopher F Basler Reed S Shabman |
| author_facet | Kari A Dilley Alexander A Voorhies Priya Luthra Vinita Puri Timothy B Stockwell Hernan Lorenzi Christopher F Basler Reed S Shabman |
| author_sort | Kari A Dilley |
| collection | DOAJ |
| description | Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN) response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV), and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA) or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI) RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV). Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR) activation. |
| first_indexed | 2024-12-16T08:57:21Z |
| format | Article |
| id | doaj.art-03bbf0a609424c4caa0200b46b760f9b |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-16T08:57:21Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-03bbf0a609424c4caa0200b46b760f9b2022-12-21T22:37:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01126e017871710.1371/journal.pone.0178717The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing.Kari A DilleyAlexander A VoorhiesPriya LuthraVinita PuriTimothy B StockwellHernan LorenziChristopher F BaslerReed S ShabmanEbola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN) response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV), and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA) or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI) RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV). Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR) activation.https://doi.org/10.1371/journal.pone.0178717 |
| spellingShingle | Kari A Dilley Alexander A Voorhies Priya Luthra Vinita Puri Timothy B Stockwell Hernan Lorenzi Christopher F Basler Reed S Shabman The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. PLoS ONE |
| title | The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. |
| title_full | The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. |
| title_fullStr | The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. |
| title_full_unstemmed | The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. |
| title_short | The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing. |
| title_sort | ebola virus vp35 protein binds viral immunostimulatory and host rnas identified through deep sequencing |
| url | https://doi.org/10.1371/journal.pone.0178717 |
| work_keys_str_mv | AT kariadilley theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT alexanderavoorhies theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT priyaluthra theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT vinitapuri theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT timothybstockwell theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT hernanlorenzi theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT christopherfbasler theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT reedsshabman theebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT kariadilley ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT alexanderavoorhies ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT priyaluthra ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT vinitapuri ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT timothybstockwell ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT hernanlorenzi ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT christopherfbasler ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing AT reedsshabman ebolavirusvp35proteinbindsviralimmunostimulatoryandhostrnasidentifiedthroughdeepsequencing |