Non-canonical proline-tyrosine interactions with multiple host proteins regulate Ebola virus infection

Jyoti Batra, Hiroyuki Mori, Gabriel I. Small, Manu Anantpadma, Olena Shtanko, Nawneet Mishra, Mengru Zhang, Dandan Liu, Caroline G. Williams, Nadine Biedenkopf, Stephan Becker, Michael L. Gross, Daisy W. Leung, Robert A. Davey, Gaya K. Amarasinghe, Nevan J. Krogan, Christopher F. Basler

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The Ebola virus VP30 protein interacts with the viral nucleoprotein and with host protein RBBP6 via PPxPxY motifs that adopt non-canonical orientations, as compared to other proline-rich motifs. An affinity tag-purification mass spectrometry approach identified additional PPxPxY-containing host proteins hnRNP L, hnRNPUL1, and PEG10, as VP30 interactors. hnRNP L and PEG10, like RBBP6, inhibit viral RNA synthesis and EBOV infection, whereas hnRNPUL1 enhances. RBBP6 and hnRNP L modulate VP30 phosphorylation, increase viral transcription, and exert additive effects on viral RNA synthesis. PEG10 has more modest inhibitory effects on EBOV replication. hnRNPUL1 positively affects viral RNA synthesis but in a VP30-independent manner. Binding studies demonstrate variable capacity of the PPxPxY motifs from these proteins to bind VP30, define PxPPPPxY as an optimal binding motif, and identify the fifth proline and the tyrosine as most critical for interaction. Competition binding and hydrogen-deuterium exchange mass spectrometry studies demonstrate that each protein binds a similar interface on VP30. VP30 therefore presents a novel proline recognition domain that is targeted by multiple host proteins to modulate viral transcription.

Original languageEnglish
Article numbere105658
JournalEMBO Journal
Volume40
Issue number18
DOIs
StatePublished - Sep 15 2021

Keywords

  • Ebola virus
  • RNA viruses
  • VP30
  • viral replication
  • virus–host interactions

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