Targeting Membrane-Bound Viral RNA Synthesis Reveals Potent Inhibition of Diverse Coronaviruses Including the Middle East Respiratory Syndrome Virus
Journal article, 2014
Coronaviruses raise serious concerns as emerging zoonotic viruses without specific antiviral drugs available. Here we screened a collection of 16671 diverse compounds for anti-human coronavirus 229E activity and identified an inhibitor, designated K22, that specifically targets membrane-bound coronaviral RNA synthesis. K22 exerts most potent antiviral activity after virus entry during an early step of the viral life cycle. Specifically, the formation of double membrane vesicles (DMVs), a hallmark of coronavirus replication, was greatly impaired upon K22 treatment accompanied by near-complete inhibition of viral RNA synthesis. K22-resistant viruses contained substitutions in non-structural protein 6 (nsp6), a membrane-spanning integral component of the viral replication complex implicated in DMV formation, corroborating that K22 targets membrane bound viral RNA synthesis. Besides K22 resistance, the nsp6 mutants induced a reduced number of DMVs, displayed decreased specific infectivity, while RNA synthesis was not affected. Importantly, K22 inhibits a broad range of coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV), and efficient inhibition was achieved in primary human epithelia cultures representing the entry port of human coronavirus infection. Collectively, this study proposes an evolutionary conserved step in the life cycle of positive-stranded RNA viruses, the recruitment of cellular membranes for viral replication, as vulnerable and, most importantly, druggable target for antiviral intervention. We expect this mode of action to serve as a paradigm for the development of potent antiviral drugs to combat many animal and human virus infections.
Parasitology
3C-LIKE
REPLICATION COMPLEX
Microbiology
MAIN PROTEINASE
VACCINIA VIRUS
Virology
CELL-CULTURES
DOUBLE-STRANDED-RNA
IN-VITRO
FUNCTIONAL RECEPTOR
MOUSE HEPATITIS-VIRUS
SARS CORONAVIRUS
PROTEINASE
Author
Anna Lundin
University of Gothenburg
R. Dijkman
Federal Department of Home Affairs
Kantonsspital St Gallen
Tomas Bergström
University of Gothenburg
Nina Kann
Chalmers, Chemical and Biological Engineering, Organic Chemistry
Beata Adamiak
University of Gothenburg
Charles Hannoun
University of Gothenburg
E. Kindler
Kantonsspital St Gallen
Federal Department of Home Affairs
H. R. Jonsdottir
Kantonsspital St Gallen
Federal Department of Home Affairs
D. Muth
Universitat Bonn
J. Kint
Wageningen University and Research Centre
Merck Animal Health
M. Forlenza
Wageningen University and Research Centre
M. A. Muller
Universitat Bonn
C. Drosten
Universitat Bonn
V. Thiel
Kantonsspital St Gallen
Universitat Bern
Federal Department of Home Affairs
Edward Trybala
University of Gothenburg
PLoS Pathogens
1553-7366 (ISSN) 1553-7374 (eISSN)
Vol. 10 5 e1004166- e1004166Subject Categories (SSIF 2011)
Microbiology
DOI
10.1371/journal.ppat.1004166