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Visualizing Herpesvirus Procapsids in Living Cells.

Oana Maier1, Patricia J Sollars2, Gary E Pickard2

  • 1Department of Microbiology-Immunology, Northwestern University, Chicago, Illinois, USA.

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|September 2, 2016
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Summary
This summary is machine-generated.

Researchers developed new fluorescent herpesvirus (human and veterinary pathogens) to image capsid assembly in living cells. This breakthrough allows visualization of procapsids and monitoring of maturation dynamics, offering new insights into viral infection processes.

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Area of Science:

  • Virology
  • Cell Biology
  • Microbiology

Background:

  • Herpesviridae are significant human and veterinary pathogens.
  • Understanding herpesvirus capsid assembly and maturation is crucial but challenging in live cells.
  • Existing fluorescent tags do not label initial capsid structures (procapsids).

Purpose of the Study:

  • To develop a method for live-cell imaging of herpesvirus procapsids.
  • To investigate the dynamics of capsid assembly and maturation in real-time.
  • To gain new insights into herpesvirus morphogenesis.

Main Methods:

  • Created recombinant pseudorabies viruses encoding green fluorescent protein (GFP) fused to internal capsid proteins.
  • Utilized a GFP-VP24 fusion protein as a procapsid marker.
  • Employed dual-fluorescent viruses (GFP-VP24 and pUL25/mCherry) for simultaneous imaging.
  • Assessed viral propagation kinetics and virulence in vitro and in vivo.

Main Results:

  • A GFP-VP24 fusion protein was successfully created and tolerated in herpes simplex virus.
  • GFP-VP24 served as a reliable marker for procapsids, remaining associated with the capsid.
  • Dual-fluorescent viruses enabled discrimination of procapsids and monitoring of capsid maturation.
  • Confirmed the feasibility of imaging herpesvirus procapsids and their morphogenesis in living cells.

Conclusions:

  • The study demonstrates the successful development of tools for live-cell imaging of herpesvirus procapsids.
  • Herpesvirus capsid shell maturation kinetics can now be monitored in real-time.
  • The encapsidation machinery appears to play a limited role in coordinating capsid shell maturation.