Ubiquitin (Ub) is a vital regulatory component in various cellular processes, including cellular responses to
viral infection. As obligate intracellular pathogens, viruses have the capacity to manipulate the
ubiquitin (Ub) cycle to their advantage by encoding Ub-modifying
proteins including
deubiquitinases (DUBs). However, how cellular DUBs modulate specific
viral infections, such as norovirus, is poorly understood. To examine the role of DUBs during
norovirus infection, we used
WP1130, a small molecule inhibitor of a subset of cellular DUBs. Replication of murine norovirus in murine macrophages and the human norovirus Norwalk virus in a replicon system were significantly inhibited by
WP1130. Chemical proteomics identified the cellular DUB USP14 as a target of
WP1130 in murine macrophages, and pharmacologic inhibition or
siRNA-mediated knockdown of USP14 inhibited murine
norovirus infection. USP14 is a
proteasome-associated DUB that also binds to
inositol-requiring
enzyme 1 (IRE1), a critical mediator of the unfolded protein response (UPR).
WP1130 treatment of murine macrophages did not alter
proteasome activity but activated the X-box binding protein-1 (XBP-1) through an IRE1-dependent mechanism. In addition,
WP1130 treatment or induction of the UPR also reduced
infection of other RNA viruses including encephalomyocarditis virus, Sindbis virus, and La Crosse virus but not
vesicular stomatitis virus. Pharmacologic inhibition of the IRE1
endonuclease activity partially rescued the
antiviral effect of
WP1130. Taken together, our studies support a model whereby induction of the UPR through cellular DUB inhibition blocks specific
viral infections, and suggest that cellular DUBs and the UPR represent novel targets for future development of broad spectrum
antiviral therapies.