Human cytomegalovirus (HCMV) is an important pathogen with multiple immune evasion strategies, including virally facilitated degradation of host antiviral restriction factors. Here, we describe a multiplexed approach to discover proteins with innate immune function on the basis of active degradation by the proteasome or lysosome during early-phase HCMV infection. Using three orthogonal proteomic/transcriptomic screens to quantify protein degradation, with high confidence we identified 35 proteins enriched in antiviral restriction factors. A final screen employed a comprehensive panel of viral mutants to predict viral genes that target >250 human proteins. This approach revealed that helicase-like transcription factor (HLTF), a DNA helicase important in DNA repair, potently inhibits early viral gene expression but is rapidly degraded during infection. The functionally unknown HCMV protein UL145 facilitates HLTF degradation by recruiting the Cullin4 E3 ligase complex. Our approach and data will enable further identifications of innate pathways targeted by HCMV and other viruses.

• Klíčová slova
• host-pathogen interaction, immune evasion, innate immunity, lysosome, proteasome, protein degradation, pulsed SILAC, quantitative proteomics, restriction factor, tandem mass tag,
• MeSH
• cytomegalovirové infekce genetika   imunologie   virologie   MeSH
• Cytomegalovirus genetika   imunologie   fyziologie   MeSH
• DNA vazebné proteiny chemie   genetika   imunologie   MeSH
• imunitní únik MeSH
• lidé MeSH
• proteiny chemie   genetika   imunologie   MeSH
• proteomika MeSH
• stabilita proteinů MeSH
• transkripční faktory chemie   genetika   imunologie   MeSH
• virové proteiny chemie   genetika   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA vazebné proteiny MeSH
• HLTF protein, human MeSH Prohlížeč
• proteiny MeSH
• transkripční faktory MeSH
• virové proteiny MeSH