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

Jyoti Batra; Judd F Hultquist; Dandan Liu; Olena Shtanko; John Von Dollen; Laura Satkamp; Gwendolyn M Jang; Priya Luthra; Toni M Schwarz; Gabriel I Small; Eusondia Arnett; Manu Anantpadma; Ann Reyes; Daisy W Leung; Robyn Kaake; Paige Haas; Carson B Schmidt; Larry S Schlesinger; Douglas J LaCount; Robert A Davey; Gaya K Amarasinghe; Christopher F Basler; Nevan J Krogan

doi:10.1016/j.cell.2018.08.044

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

Cell. 2018 Dec 13;175(7):1917-1930.e13. doi: 10.1016/j.cell.2018.08.044.

Authors

Jyoti Batra¹, Judd F Hultquist², Dandan Liu³, Olena Shtanko⁴, John Von Dollen⁵, Laura Satkamp⁵, Gwendolyn M Jang⁵, Priya Luthra⁶, Toni M Schwarz⁷, Gabriel I Small³, Eusondia Arnett⁸, Manu Anantpadma⁴, Ann Reyes⁴, Daisy W Leung³, Robyn Kaake⁵, Paige Haas⁵, Carson B Schmidt⁵, Larry S Schlesinger⁸, Douglas J LaCount⁹, Robert A Davey⁴, Gaya K Amarasinghe³, Christopher F Basler¹⁰, Nevan J Krogan¹¹

Affiliations

¹ Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA; Quantitative Biosciences Institute, University of California, San Francisco, San Francisco, CA 94158, USA; J. David Gladstone Institutes, San Francisco, CA 94158, USA.
² Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Quantitative Biosciences Institute, University of California, San Francisco, San Francisco, CA 94158, USA; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Division of Infectious Diseases, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
³ Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63105, USA.
⁴ Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX 78245, USA.
⁵ Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Quantitative Biosciences Institute, University of California, San Francisco, San Francisco, CA 94158, USA; J. David Gladstone Institutes, San Francisco, CA 94158, USA.
⁶ Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA.
⁷ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁸ Texas Biomedical Research Institute, San Antonio, TX 78245, USA.
⁹ Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA.
¹⁰ Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA. Electronic address: cbasler@gsu.edu.
¹¹ Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Quantitative Biosciences Institute, University of California, San Francisco, San Francisco, CA 94158, USA; J. David Gladstone Institutes, San Francisco, CA 94158, USA. Electronic address: nevan.krogan@ucsf.edu.

Abstract

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.

Keywords: Ebola virus; RBBP6; RNA viruses; VP30; antiviral factor; host-pathogen interactions; protein-protein interactions; virus-host interactions.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Carrier Proteins* / chemistry
Carrier Proteins* / genetics
Carrier Proteins* / metabolism
Crystallography, X-Ray
DNA-Binding Proteins* / chemistry
DNA-Binding Proteins* / genetics
DNA-Binding Proteins* / metabolism
Ebolavirus / physiology*
HEK293 Cells
HeLa Cells
Hemorrhagic Fever, Ebola / genetics
Hemorrhagic Fever, Ebola / metabolism*
Hemorrhagic Fever, Ebola / pathology
Humans
Protein Interaction Mapping
Transcription Factors* / chemistry
Transcription Factors* / genetics
Transcription Factors* / metabolism
Ubiquitin-Protein Ligases
Viral Proteins* / chemistry
Viral Proteins* / genetics
Viral Proteins* / metabolism
Virus Replication / physiology*

Substances

Carrier Proteins
DNA-Binding Proteins
Transcription Factors
VP30 protein, ebola virus
Viral Proteins
RBBP6 protein, human
Ubiquitin-Protein Ligases

Abstract

Publication types

MeSH terms

Substances

Grants and funding