Ubiquitin variants potently inhibit SARS-CoV-2 PLpro and viral replication via a novel site distal to the protease active site

Vera J.E. van Vliet, Nhan Huynh, Judith Palà, Ankoor Patel, Alex Singer, Cole Slater, Jacky Chung, Mariska van Huizen, Joan Teyra, Shane Miersch, Gia Khanh Luu, Wei Ye, Nitin Sharma, Safder S. Ganaie, Raquel Russell, Chao Chen, Mindy Maynard, Gaya K. Amarasinghe, Brian L. Mark, Marjolein KikkertSachdev S. Sidhu

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has made it clear that combating coronavirus outbreaks benefits from a combination of vaccines and therapeutics. A promising drug target common to all coronaviruses—including SARS-CoV, MERS-CoV, and SARS-CoV-2—is the papain-like protease (PLpro). PLpro cleaves part of the viral replicase polyproteins into non-structural protein subunits, which are essential to the viral replication cycle. Additionally, PLpro can cleave both ubiquitin and the ubiquitin-like protein ISG15 from host cell substrates as a mechanism to evade innate immune responses during infection. These roles make PLpro an attractive antiviral drug target. Here we demonstrate that ubiquitin variants (UbVs) can be selected from a phage-displayed library and used to specifically and potently block SARS-CoV-2 PLpro activity. A crystal structure of SARS-CoV-2 PLpro in complex with a representative UbV reveals a dimeric UbV bound to PLpro at a site distal to the catalytic site. Yet, the UbV inhibits the essential cleavage activities of the protease in vitro and in cells, and it reduces viral replication in cell culture by almost five orders of magnitude.

Original languageEnglish
Article numbere1011065
JournalPLoS pathogens
Volume18
Issue number12
DOIs
StatePublished - Dec 2022

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