Proteotype profiling unmasks a viral signalling network essential for poxvirus assembly and transcriptional competence

Research output: Contribution to journalArticle

Authors

  • Karel Novy
  • Samuel Kilcher
  • Ulrich Omasits
  • Christopher Karl Ernst Bleck
  • Corina Beerli
  • Jakob Vowinckel
  • Caroline K. Martin
  • Mohammedyaseen Syedbasha
  • Alessio Maiolica
  • Ian White
  • Bernd Wollscheid

Colleges, School and Institutes

External organisations

  • HPK H27
  • UCL
  • Biozentrum, Universität Basel
  • Biognosys AG

Abstract

To orchestrate context-dependent signalling programmes, poxviruses encode two dual-specificity enzymes, the F10 kinase and the H1 phosphatase. These signalling mediators are essential for poxvirus production, yet their substrate profiles and systems-level functions remain enigmatic. Using a phosphoproteomic screen of cells infected with wild-type, F10 and H1 mutant vaccinia viruses, we systematically defined the viral signalling network controlled by these enzymes. Quantitative cross-comparison revealed 33 F10 and/or H1 phosphosites within 17 viral proteins. Using this proteotype dataset to inform genotype-phenotype relationships, we found that H1-deficient virions harbour a hidden hypercleavage phenotype driven by reversible phosphorylation of the virus protease I7 (S134). Quantitative phosphoproteomic profiling further revealed that the phosphorylation-dependent activity of the viral early transcription factor, A7 (Y367), underlies the transcription-deficient phenotype of H1 mutant virions. Together, these results highlight the utility of combining quantitative proteotype screens with mutant viruses to uncover proteotype-phenotype-genotype relationships that are masked by classical genetic studies.

Details

Original languageEnglish
Pages (from-to)588-599
Number of pages12
JournalNature Microbiology
Volume3
Issue number5
Early online date9 Apr 2018
Publication statusPublished - May 2018