Viruses, Vol. 12, Pages 105: Reporter Assays for Ebola Virus Nucleoprotein Oligomerization, Virion-Like Particle Budding, and Minigenome Activity Reveal the Importance of Nucleoprotein Amino Acid Position 111
Viruses, Vol. 12, Pages 105: Reporter Assays for Ebola Virus Nucleoprotein Oligomerization, Virion-Like Particle Budding, and Minigenome Activity Reveal the Importance of Nucleoprotein Amino Acid Position 111
Viruses doi: 10.3390/v12010105
Authors:
Lin
Diehl
Cai
Finch
Akusobi
Kirchdoerfer
Bollinger
Schaffner
Brown
Saphire
Andersen
Kuhn
Luban
Sabeti
For highly pathogenic viruses, reporter assays that can be rapidly performed are critically needed to identify potentially functional mutations for further study under maximal containment (e.g., biosafety level 4 [BSL-4]). The Ebola virus nucleoprotein (NP) plays multiple essential roles during the viral life cycle, yet few tools exist to study the protein under BSL-2 or equivalent containment. Therefore, we adapted reporter assays to measure NP oligomerization and virion-like particle (VLP) production in live cells and further measured transcription and replication using established minigenome assays. As a proof-of-concept, we examined the NP-R111C substitution, which emerged during the 2013‒2016 Western African Ebola virus disease epidemic and rose to high frequency. NP-R111C slightly increased NP oligomerization and VLP budding but slightly decreased transcription and replication. By contrast, a synthetic charge-reversal mutant, NP-R111E, greatly increased oligomerization but abrogated transcription and replication. These results are intriguing in light of recent structures of NP...
Source: Viruses - Category: Virology Authors: Lin Diehl Cai Finch Akusobi Kirchdoerfer Bollinger Schaffner Brown Saphire Andersen Kuhn Luban Sabeti Tags: Article Source Type: research