An Intrinsically Disordered Peptide from Ebola Virus VP35 Controls Viral RNA Synthesis by Modulating Nucleoprotein-RNA Interactions
Daisy W. Leung,
Dominika Borek,
Priya Luthra,
Jennifer M. Binning,
Manu Anantpadma,
Gai Liu,
Ian B. Harvey,
Zhaoming Su,
Ariel Endlich-Frazier,
Juanli Pan,
Reed S. Shabman,
Wah Chiu,
Robert A. Davey,
Zbyszek Otwinowski,
Christopher F. Basler,
Gaya K. Amarasinghe
Affiliations
Daisy W. Leung
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
Dominika Borek
Departments of Biophysics and Biochemistry and Center for Structural Genomics of Infectious Diseases, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390, USA
Priya Luthra
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Jennifer M. Binning
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
Manu Anantpadma
Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX 78227, USA
Gai Liu
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
Ian B. Harvey
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
Zhaoming Su
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA
Ariel Endlich-Frazier
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Juanli Pan
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
Reed S. Shabman
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Wah Chiu
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA
Robert A. Davey
Department of Virology and Immunology, Texas Biomedical Research Institute, San Antonio, TX 78227, USA
Zbyszek Otwinowski
Departments of Biophysics and Biochemistry and Center for Structural Genomics of Infectious Diseases, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390, USA
Christopher F. Basler
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Gaya K. Amarasinghe
Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO 63110, USA
During viral RNA synthesis, Ebola virus (EBOV) nucleoprotein (NP) alternates between an RNA-template-bound form and a template-free form to provide the viral polymerase access to the RNA template. In addition, newly synthesized NP must be prevented from indiscriminately binding to noncognate RNAs. Here, we investigate the molecular bases for these critical processes. We identify an intrinsically disordered peptide derived from EBOV VP35 (NPBP, residues 20–48) that binds NP with high affinity and specificity, inhibits NP oligomerization, and releases RNA from NP-RNA complexes in vitro. The structure of the NPBP/ΔNPNTD complex, solved to 3.7 Å resolution, reveals how NPBP peptide occludes a large surface area that is important for NP-NP and NP-RNA interactions and for viral RNA synthesis. Together, our results identify a highly conserved viral interface that is important for EBOV replication and can be targeted for therapeutic development.
