Discovery of small-molecule inhibitors targeting the E3 ubiquitin ligase activity of the herpes simplex virus 1 ICP0 protein using an in vitro high-throughput screening assay

Thibaut Deschamps, Hope Waisner, Christos Dogrammatzis, Anuradha Roy, Shibin Chacko, Chamani Perera, Thomas E. Prisinzano, Maria Kalamvoki

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Herpes simplex virus 1 (HSV-1) has infected more than 80% of the population. Reactivation of the virus causes diseases ranging in severity from benign cold sores to fatal encephalitis. Current treatments involve viral DNA replication inhibitors, but the emergence of drug-resistant mutants is observed frequently, highlighting the need for novel antiviral therapies. Infected cell protein 0 (ICP0) of HSV-1 is encoded by an immediate early gene and plays a fundamental role during infection, because it enables viral gene expression and blocks antiviral responses. One mechanism by which ICP0 functions is through an E3 ubiquitin ligase activity that induces the degradation of targeted proteins. A ΔICP0 virus or mutants with deficiencies in E3 ligase activity cannot counteract beta interferon (IFN-)-induced restriction of viral infection, are highly immunogenic, are avirulent, and fail to spread. Thus, small molecules interfering with essential and conserved ICP0 functions are expected to compromise HSV-1 infection. We have developed a high-throughput screening assay, based on the autoubiquitination properties of ICP0, to identify small-molecule inhibitors of ICP0 E3 ubiquitin ligase activity. Through a pilot screening procedure, we identified nine compounds that displayed dose-dependent inhibitory effects on ICP0 but not on Mdm2, a control E3 ubiquitin ligase. Following validation, one compound displayed ICP0-dependent inhibition of HSV-1 infection. This compound appeared to bind ICP0 in a cellular thermal shift assay, it blocked ICP0 self-elimination, and it blocked wild-type but not ICP0-null virus gene expression. This scaffold displays specificity and could be used to develop optimized ICP0 E3 ligase inhibitors.

Original languageEnglish
Article numbere00619-19
JournalJournal of Virology
Volume93
Issue number13
DOIs
StatePublished - 2019

Bibliographical note

Publisher Copyright:
© 2019 American Society for Microbiology. All Rights Reserved.

Keywords

  • E3 ubiquitin ligase
  • HSV
  • High-throughput screening assay
  • ICP0
  • Small-molecule inhibitors

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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