Project description:Primary human foreskin fibroblasts (HFF) were infected with wild-type herpes simplex virus 1 (HSV-1) strain 17 or its vhs null mutant, or with BAC-derived viruses expressing either a wild-type or catalytically inactive vhs with the D195N mutation at a multiplicity of infection (MOI) of 10. Chromatin-associated RNA was prepared and subjected to RNA-seq.
Project description:Human herpesviruses are widespread human pathogens with a remarkable impact on worldwide public health. Despite intense decades of research, the molecular details in many aspects of their function remain to be fully characterized. To unravel the details of how these viruses operate a thorough understanding of the relationships between the viral protein components is key. Here, we have created HVint, a novel protein-protein intra-viral interaction database for herpes simplex virus type 1 (HSV-1), which integrates data from five external sources. The coverage of the initial interactome was enlarged using evolutionary information, which integrates computational predictions for potential novel interactions in HSV-1. As an experimental validation for these predicted interactions, affinity purification mass spectrometry (AP-MS) was performed for the HSV-1 capsid protein, VP26. The VP26 interaction data provided experimental evidence supporting the interactions with pUL31 and pUL40, which were computationally predicted as direct partners. Other indirect interactions within the predicted network, such as pUL32 and pUL21, were also present in the AP-MS experiment. For access to the complete HVint network, we have developed a user-friendly and interactive web interface. Our approach demonstrates the power of computational predictions to assist in the design of targeted experiments for the discovery of novel protein-protein interactions.