TY - JOUR
T1 - Proteotype profiling unmasks a viral signalling network essential for poxvirus assembly and transcriptional competence
AU - Novy, Karel
AU - Kilcher, Samuel
AU - Omasits, Ulrich
AU - Bleck, Christopher Karl Ernst
AU - Beerli, Corina
AU - Vowinckel, Jakob
AU - Martin, Caroline K.
AU - Syedbasha, Mohammedyaseen
AU - Maiolica, Alessio
AU - White, Ian
AU - Mercer, Jason
AU - Wollscheid, Bernd
PY - 2018/5
Y1 - 2018/5
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85045091791&partnerID=8YFLogxK
U2 - 10.1038/s41564-018-0142-6
DO - 10.1038/s41564-018-0142-6
M3 - Article
C2 - 29632367
AN - SCOPUS:85045091791
SN - 2058-5276
VL - 3
SP - 588
EP - 599
JO - Nature Microbiology
JF - Nature Microbiology
IS - 5
ER -