Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

• Published in: Cell Vol. 175; no. 7; pp. 1917 - 1930.e13
• Main Authors: Batra, Jyoti, Hultquist, Judd F., Liu, Dandan, Shtanko, Olena, Von Dollen, John, Satkamp, Laura, Jang, Gwendolyn M., Luthra, Priya, Schwarz, Toni M., Small, Gabriel I., Arnett, Eusondia, Anantpadma, Manu, Reyes, Ann, Leung, Daisy W., Kaake, Robyn, Haas, Paige, Schmidt, Carson B., Schlesinger, Larry S., LaCount, Douglas J., Davey, Robert A., Amarasinghe, Gaya K., Basler, Christopher F., Krogan, Nevan J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 13.12.2018
• Subjects: antiviral factor; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - metabolism; Crystallography, X-Ray; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Ebola virus; Ebolavirus - physiology; HEK293 Cells; HeLa Cells; Hemorrhagic Fever, Ebola - genetics; Hemorrhagic Fever, Ebola - metabolism; Hemorrhagic Fever, Ebola - pathology; host-pathogen interactions; Humans; Protein Interaction Mapping; protein-protein interactions; RBBP6; RNA viruses; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - metabolism; Ubiquitin-Protein Ligases; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - metabolism; Virus Replication - physiology; virus-host interactions; VP30; Ebola virus; antiviral factor; RNA viruses; host-pathogen interactions; virus-host interactions; RBBP6; protein-protein interactions; VP30
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/j.cell.2018.08.044

Ebola virus (EBOV) infection often results in fatal illness in humans, yet little is known about how EBOV usurps host pathways during infection. To address this, we used affinity tag-purification mass spectrometry (AP-MS) to generate an EBOV-host protein-protein interaction (PPI) map. We uncovered 194 high-confidence EBOV-human PPIs, including one between the viral transcription regulator VP30 and the host ubiquitin ligase RBBP6. Domain mapping identified a 23 amino acid region within RBBP6 that binds to VP30. A crystal structure of the VP30-RBBP6 peptide complex revealed that RBBP6 mimics the viral nucleoprotein (NP) binding to the same interface of VP30. Knockdown of endogenous RBBP6 stimulated viral transcription and increased EBOV replication, whereas overexpression of either RBBP6 or the peptide strongly inhibited both. These results demonstrate the therapeutic potential of biologics that target this interface and identify additional PPIs that may be leveraged for novel therapeutic strategies. [Display omitted] •194 high-confidence Ebola virus-host protein interactions identified by AP-MS•Host protein RBBP6 targets the VP30 protein at the viral nucleoprotein binding cleft•RBBP6 competes with NP for binding to VP30 and inhibits Ebola virus RNA synthesis•RBBP6-derived peptides potently inhibit Ebola virus infection A host-Ebola protein-protein interaction map identifies candidate drug targets, including a host ubiquitin ligase that prevents viral replication, peptide mimics of which strongly inhibit Ebola replication in multiple cell types.