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Dembowski, J. A.

Publications and source records attributed to Dembowski, J. A..

2 recordsLinked to original sources

Distinct labeling of host and viral genomes highlights redistribution of nuclear proteins during HSV-1 infection

Herpes simplex virus type 1 (HSV-1) coordinates viral and cellular proteins during infection. While proteins associated with HSV-1 genomes have been identified, changes in proteins associated with the host genome during infection remain poorly understood. We adapted accelerated native isolation of proteins on nascent DNA (aniPOND) to label and purify cellular and viral DNA then identify associated proteins by mass spectrometry. Using this approach, we confirm observations that cellular transcription, DNA replication, and repair proteins are enriched on viral genomes after the onset of viral DNA replication. We assessed how protein interactions with cellular DNA change during infection relative to protein abundance. Targets of the viral E3 ubiquitin ligase, ICP0, are depleted from the nucleus and cellular genome during infection. Transcription regulatory factors, Pol II, and the proteasome are depleted from the host genome, while association with DNA damage response and nuclear pore proteins increases. We also provide evidence for HSV-1 disruption of ribosome biogenesis. Results demonstrate reorganization of the nucleus during infection and validate this technique to identify changes to host genome-associated factors during infection.

molecular biology↗

Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection

Herpesviruses utilize multiple mechanisms to redirect host proteins for use in viral processes and to avoid recognition and repression by the host. To investigate the dynamic interactions between HSV-1 DNA and viral and host proteins, we developed an approach to identify proteins that associate with the infecting viral genome from nuclear entry through packaging. We found that input viral DNA progressed within six hours through four temporal stages where the genomes: 1. interacted with intrinsic and DNA damage response proteins, 2. underwent a robust transcriptional switch mediated largely by ICP4, 3. engaged in replication, repair, and continued transcription, and then 4. transitioned to a more transcriptionally inert state engaging de novo synthesized viral structural components while maintaining interactions with replication proteins. Using a combination of genetic, imaging, and proteomic approaches, we provide a new and temporally compressed view of the HSV-1 life cycle based on genome-proteome dynamics.

microbiology↗