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Updated: Dec 28, 2025
Temporal Analysis of the Nuclear-to-cytoplasmic Translocation of a Herpes Simplex Virus 1 Protein by Immunofluorescent Confocal Microscopy
Published on: November 4, 2018
Temporal dynamics of protein complex formation and dissociation during human cytomegalovirus infection
Yutaka Hashimoto1,2,3,4, Xinlei Sheng1, Laura A Murray-Nerger1
1Department of Molecular Biology, Princeton University, Lewis Thomas Laboratory, Washington Road, Princeton, NJ, 08544, USA.
Abstract:
The co-evolution and co-existence of viral pathogens with their hosts for millions of years is reflected in dynamic virus-host protein-protein interactions (PPIs) that are intrinsic to the spread of infections. Here, we investigate the system-wide dynamics of protein complexes throughout infection with the herpesvirus, human cytomegalovirus (HCMV). Integrating thermal shift assays and mass spectrometry quantification with virology and microscopy, we monitor the temporal formation and dissociation of hundreds of functional protein complexes and the dynamics of host-host, virus-host, and virus-virus PPIs. We establish pro-viral roles for cellular protein complexes and translocating proteins. We show the HCMV receptor integrin beta 1 dissociates from extracellular matrix proteins, becoming internalized with CD63, which is necessary for virus production. Moreover, this approach facilitates characterization of essential viral proteins, such as pUL52. This study of temporal protein complex dynamics provides insights into mechanisms of HCMV infection and a resource for biological and therapeutic studies.
Insights
Human cytomegalovirus (HCMV) infection involves dynamic protein interactions. This study maps these interactions over time, revealing key viral and host protein roles in infection spread and essential viral proteins.
Area of Science:
- Virology
- Molecular Biology
- Systems Biology
Background:
- Viral pathogens and hosts co-evolve, leading to complex protein-protein interactions (PPIs) crucial for infection.
- Understanding these dynamic interactions is key to deciphering viral spread and developing therapies.
Purpose of the Study:
- To investigate the system-wide dynamics of protein complexes during human cytomegalovirus (HCMV) infection.
- To identify the temporal formation and dissociation of host-host, virus-host, and virus-virus PPIs.
Main Methods:
- Integration of thermal shift assays, mass spectrometry, virology, and microscopy.
- Monitoring of hundreds of functional protein complexes and PPI dynamics throughout the infection cycle.
Main Results:
- Pro-viral roles were established for cellular protein complexes and translocating proteins.
- HCMV receptor integrin beta 1 dissociation and internalization with CD63 were shown to be essential for virus production.
- Essential viral proteins, such as pUL52, were characterized.
Conclusions:
- The study provides insights into the mechanisms of HCMV infection through temporal protein complex dynamics.
- This work serves as a valuable resource for future biological and therapeutic studies on HCMV.
