Herpes simplex virus 1 infection induces ubiquitination of UBE1a

Biochem J. 2021 Jan 15;478(1):261-279. doi: 10.1042/BCJ20200885.

Abstract

Herpes simplex virus 1 (HSV-1) is a human DNA virus that causes cold sores, keratitis, meningitis, and encephalitis. Ubiquitination is a post-translational protein modification essential for regulation of cellular events, such as proteasomal degradation, signal transduction, and protein trafficking. The process is also involved in events for establishing viral infection and replication. The first step in ubiquitination involves ubiquitin (Ub) binding with Ub-activating enzyme (E1, also termed UBE1) via a thioester linkage. Our results show that HSV-1 infection alters protein ubiquitination pattern in host cells, as evidenced by MS spectra and co-immunoprecipitation assays. HSV-1 induced ubiquitination of UBE1a isoform via an isopeptide bond with Lys604. Moreover, we show that ubiquitination of K604 in UBE1a enhances UBE1a activity; that is, the activity of ubiquitin-transfer to E2 enzyme. Subsequently, we investigated the functional role of UBE1a and ubiquitination of K604 in UBE1a. We found that UBE1-knockdown increased HSV-1 DNA replication and viral production. Furthermore, overexpression of UBE1a, but not a UBE1a K604A mutant, suppressed viral replication. Furthermore, we found that UBE1a and ubiquitination at K604 in UBE1a retarded expression of HSV-1 major capsid protein, ICP5. Our findings show that UBE1a functions as an antiviral factor that becomes activated upon ubiquitination at Lys604.

Keywords: UBE1; herpesvirus; post-translational modification; ubiquitin proteasome system; ubiquitins; virus replication.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / metabolism*
  • Cell Survival / genetics
  • Chlorocebus aethiops
  • Chromatography, Liquid
  • Doxycycline / pharmacology
  • HeLa Cells
  • Herpes Simplex / enzymology
  • Herpes Simplex / genetics
  • Herpes Simplex / metabolism*
  • Herpesvirus 1, Human / genetics
  • Herpesvirus 1, Human / metabolism*
  • Host-Pathogen Interactions / drug effects*
  • Host-Pathogen Interactions / genetics
  • Humans
  • Protein Binding
  • Protein Domains
  • Protein Processing, Post-Translational / drug effects
  • RNA, Small Interfering
  • Tandem Mass Spectrometry
  • Tetracycline / pharmacology
  • Transfection
  • Ubiquitin-Activating Enzymes / genetics
  • Ubiquitin-Activating Enzymes / metabolism*
  • Ubiquitination / drug effects
  • Vero Cells
  • Virus Replication / drug effects

Substances

  • Antiviral Agents
  • RNA, Small Interfering
  • UBA1 protein, human
  • Ubiquitin-Activating Enzymes
  • Tetracycline
  • Doxycycline