Literature
Potent and selective inhibition of pathogenic viruses by engineered ubiquitin variants.
Zhang, W., Bailey-Elkin, B.A., Knaap, R.C.M., Khare, B., Dalebout, T.J., Johnson, G.G., van Kasteren, P.B., McLeish, N.J., Gu, J., He, W., Kikkert, M., Mark, B.L., Sidhu, S.S.(2017) PLoS Pathog. 13: e1006372-e1006372-CoV
- PubMed: 28542609
- DOI: 10.1371/journal.ppat.1006372
- Primary Citation of Related Structures:
- PubMed Abstract:
The
recent Middle East respiratory syndrome coronavirus (MERS-CoV), Ebola
and Zika virus outbreaks exemplify the continued threat of (re-)emerging
viruses to human health, and our inability to rapidly develop effective
therapeutic countermeasures. Many viruses, including MERS-CoV and the
Crimean-Congo hemorrhagic fever virus (CCHFV) encode deubiquitinating
(DUB) enzymes that are critical for viral replication and pathogenicity.
They bind and remove ubiquitin (Ub) and interferon stimulated gene 15
(ISG15) from cellular proteins to suppress host antiviral innate immune
responses. A variety of viral DUBs (vDUBs), including the MERS-CoV
papain-like protease, are responsible for cleaving the viral replicase
polyproteins during replication, and are thereby critical components of
the viral replication cycle. Together, this makes vDUBs highly
attractive antiviral drug targets. However, structural similarity
between the catalytic cores of vDUBs and human DUBs complicates the
development of selective small molecule vDUB inhibitors. We have thus
developed an alternative strategy to target the vDUB activity through a
rational protein design approach. Here, we report the use of
phage-displayed ubiquitin variant (UbV) libraries to rapidly identify
potent and highly selective protein-based inhibitors targeting the DUB
domains of MERS-CoV and CCHFV. UbVs bound the vDUBs with high affinity
and specificity to inhibit deubiquitination, deISGylation and in the
case of MERS-CoV also viral replicative polyprotein processing.
Co-crystallization studies further revealed critical molecular
interactions between UbVs and MERS-CoV or CCHFV vDUBs, accounting for
the observed binding specificity and high affinity. Finally, expression
of UbVs during MERS-CoV infection reduced infectious progeny titers by
more than four orders of magnitude, demonstrating the remarkable potency
of UbVs as antiviral agents. Our results thereby establish a strategy
to produce protein-based inhibitors that could protect against a diverse
range of viruses by providing UbVs via mRNA or protein delivery
technologies or through transgenic techniques.
Organizational Affiliation: Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, and Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.,Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.,Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
Organizational Affiliation: Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, and Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.,Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.,Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
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