Herpesvirus virions form by remodeling of intracellular virus-host interaction networks into evolutionarily conserved particle architectures. Here, we define a virion-wide spatial and quantitative protein proximity map of herpes simplex virus 1 (HSV-1) by combining cross-linking mass spectrometry with quantitative proteomics. Integration with intracellular interaction maps reveals that maturation acts as a selective filter, transforming broad virus-host associations into an organized virion network. This process depletes biosynthetic and nuclear components while enriching interactions involved in tegument organization and envelope acquisition around the viral protein UL49. Comparison with analogous maps of human cytomegalovirus (HCMV) identifies HSV-1-UL49 and HCMV-UL32 as functionally equivalent network hubs despite lacking evolutionary relatedness. Both hubs converge on shared phosphoregulatory host factors, short linear interaction motifs, and liquid-liquid phase separation. At the virion surface, the host complement regulator CD59 protects particles from complement-mediated inactivation. Together, these findings show how conserved organizational principles shape virus-specific virion interaction networks during herpesvirus maturation.
Lars Mühlberg, Yannick Jensen, Julia Ruta et al.· bioRxiv· 0 citations
Influenza A virus (IAV) hijacks host cellular machinery during infection but many host–virus protein interactions remain uncharacterized, particularly in their native context. Here, we applied in-cell cross-linking mass spectrometry, integrated with structural modelling and functional assays, to map protein–protein contact sites in IAV-infected human cells. This revealed previously unrecognized virus–host interactions linked to spatially organized processes. We identified host factors linked to the maturation of distinct glycoforms of the viral surface glycoprotein haemagglutinin through the membrane-bound endoplasmic reticulum–Golgi system. In the nucleus, we observed the progressive disassembly of paraspeckles (phase-separated membraneless compartments) across multiple cell lines. Mechanistically, viral nucleoprotein and non-structural protein 1 interact with host paraspeckle proteins, the viral endonuclease PA-X degrades long non-coding RNA housed within paraspeckles and viral RNA polymerase II is inhibited to drive paraspeckle disruption, which releases host factors that facilitate IAV replication. These findings uncover mechanisms by which IAV exploits and remodels host compartments during infection. In-cell cross-linking mass spectrometry maps protein contacts in influenza-infected human cells, revealing how the virus hijacks host membrane-trafficking factors and dismantles nuclear paraspeckles to enhance replication.
Iuliia Kotova, Lars Mühlberg, Konstantin Gilep et al.· Nature Microbiology· 1 citation