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Simple and Robust in vivo and in vitro Approach for Studying Virus Assembly
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A Fluorescently Labeled Marburg Virus Glycoprotein as a New Tool to Study Viral Transport and Assembly

Eva Mittler1, Gordian Schudt1, Sandro Halwe1,2

  • 1Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.

The Journal of Infectious Diseases
|August 31, 2018
PubMed

Insights

Researchers developed a new Marburg virus glycoprotein (GP) tool, GP∆MLD_mCherry, for live-cell imaging. This tool successfully tracked viral protein transport and assembly, aiding in understanding filovirus infection mechanisms.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • The filovirus surface glycoprotein (GP) is crucial for cell entry and infection.
  • Understanding the intracellular trafficking and assembly of viral components is key to developing antiviral strategies.

Purpose of the Study:

  • To develop and validate a novel fluorescently tagged Marburg virus glycoprotein (GP∆MLD_mCherry) for live-cell imaging.
  • To investigate the intracellular transport and assembly pathways of Marburg virus (MARV) using the developed tool.

Main Methods:

  • Modification of Marburg virus GP by replacing the mucin-like domain with the mCherry fluorophore (GP∆MLD_mCherry).
  • Generation of a recombinant MARV expressing GP∆MLD_mCherry (recMARV MARVGP∆MLD_mCherry).
  • Live-cell imaging, including time-lapse and dual-color microscopy, to observe intracellular trafficking and protein colocalization.

Main Results:

  • GP∆MLD_mCherry exhibited similar intracellular distribution, surface transport, and virus-like particle recruitment as wild-type GP.
  • recMARV MARVGP∆MLD_mCherry-infected cells showed tubulin-dependent transport of GP∆MLD_mCherry-positive vesicles to the cell surface.
  • Dual-color imaging revealed cotransport and plasma membrane colocalization of GP∆MLD_mCherry and VP40.

Conclusions:

  • The developed GP∆MLD_mCherry is a valuable tool for studying MARV intracellular trafficking.
  • This tool facilitates the elucidation of MARV assembly pathways and provides insights into filovirus infection dynamics.

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